S. oralis - 大阪大学リポジトリ
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1.2. 18 1 2 3 4 5 6 7 8 9 mK Alefounder P R and Perham R N 1989 Identification molecular cloning and sequence analysis of a gene cluster encoding the Class II fructose 1 6 biphosphate aldolase 3 phosphoglycerate kinase and a putative second glyceraldehydes 3 phosphate dehydrogenase of Escherichia coli Mol Microbiol 3 723 732 Amano A Sojar H T Lee J Y Sharma A Levine M J and Genco R J 1994 Salivary receptors for recombinant fimbrillin of Porphyromonas gingivalis Infect Immun 62 3372 3380 Amano A Fujiwara T Nagata H Kuboniwa M Sharma A Sojar H T Genco R J Hamada S and Shizukuishi S 1997 Porphyromonas gingivalis fimbriae mediate coaggregation with Str
3. DIG GAPDH BamHI EcoRI HindlIL Kpnl PstI Sacl Sall Smal Sphl Xbal TAL GAPDH DIG High Prime DNA Labeling and Detection Starter Kit II 6 P gingivalis 381 S oralis ATCC 9811 A P gingivalis 381 S oralis ATCC 9811 FRO FEE fe B S oralis rGAPDH 2 5 pg ml OWsJW C X DEE TEL RFE REA SER 7 5S oralis GAPDH rFimA P gingivalis rFimA S oralis ATCC 9811 GAPDH BIAcore2000 GAPDH CMS E 0 05 uM 0 4 uM 5 rFimA 8 GAPDH P gingivalis 381 P gingivalis 381 14 5
4. SDS PAGE 40kDa XE 2A 2 rFimA 228 3 P gingivalis S oralis P gingivalis IFimA S oralis THES UI S oralis ERRIA P gingivalis rFimA 2B 2 P gingivalis 381 S oralis ATCC 9811 2 5 ugm 3 4 S oralis S oralis 40kDa O43 Be 6 OdJCREIESS OO N VVKVGINGFGRIGXLA ON ALQXNFXVXEGLMTTIXAYT VLDVDGK DDBJ S gordonii S pyogenes S pneumoniae S equisim
5. 10 mM PBS pH 6 0 P gingivalis rFimA GAPDH 20 ul min 37 BIAevaluation version 3 1 BIAcore 9 13 GAPDH GAPDH Pancholi 14 3 G 3 P 49 mg ml Sigma 40 mM 50 mM 5 mM EDTA pH 8 6 NAD 2 UV 1600 340 nm V NAD NADH G 3 P L GAPDH
6. 37 Az A B A x100 A B DH qQ 3 A pH GAPDH rGAPDH 25 e IE RHE 414 S oralis GAPDH GAPDHs S oralis ATCC 9811 GAPDH GAPDHs 5 3 4 N N 45 S oralis ATCC 9811 GAPDH S oralis ATCC 9811 4 ug GAPDH 10
7. X j 5 5 S oralis P gingivalis rFimA oralis DRERI N N AORO bo GAPDH CHAPS 14 Nelson S gordonii FSS2 GAPDH pH 6 5 pH 7 5 90 S oralis ATCC 9811 pH GAPDH S oralis GAPDH S oralis pH GAPDH P gingivalis rFimA rFimA P gingivalis 381 S oralis ATCC 9811
8. 0 2 um Bio Rad 15 V C45 1 10 mM PBS pH 6 0 2 000 HRP Conjugate Bio Rad 1 RRR LU 0 05 20 Tween20 PBS 15 3 HRP Conjugate Substrate Kit Bio Rad P gingivalis rFimA oralis ATCC 9811 Dyes 4 S oralis ATCC 9811 ROFEREES OT S oralis ATCC 9811 200W 1 5 000Xg 30 23 000Xg 30 1 3 3 CHAPS
9. sS oralis 10 5 S oralis ATCC 9811 GAPDH 6 P gingivalis S oralis pH GAPDH pH GAPDH Nelson S oralis ATCC 9811 pH GAPDH Bd2A 3 S oralis ATCC 9811 pH GAPDH rFimA 2B 3 p gingivalis S oralis DHE X REKEN 4 ug ml ORE CHEESE EEA SE LK 3 7 P gingivalis RESO S oralis rGAPDH S oralis ATCC 9811 GAPDH rGAPDH rGAPDH 40kDa KM 2A 4 rFimA 2B 4 P gingivali
10. 11 S oralis ATCC 9811 rGAPDH P gingivalis 381 amp 0 10 mM PBS pH 6 0 2 ml 5x108 P gingivalis 381 Sx10 5 37C 4s 3 27 S ora s ATCC 9811 oralis ATCC 10557 S gordoni i G9B sanguinis ATCC 10556 parasanguinis ATCC 15909 S oralis ATCC 9811 S oralis ATCC 10557 S gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 oralis ATCC 9811 S oralis ATCC 10557 gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 S oralis ATCC 9811 oralis ATCC 10557 S gordoni i G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 ora s ATCC 9811 oralis ATCC 10557 S gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 S oralis ATCC 9811 oralis ATCC 10557 S gordoni G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 61 61 61 61 61 121 120 121 120 120 181 180 181 180 180 241 240 241 240 240 301 300 301
11. Nyvad 4 Z S mitis S sanguinis oralis 23 V 60 90 P gingivalis GAPDH P gingivalis Zs nm P gingivalis S oralis ATCC 9811 RO SEER Ye IRE HL L 1 S oralis ATCC 9811 ROH 2 40kDa
12. gt 55 amp DIC S aureus S epidermidis GAPDH Candida albicans GAPDH GAPDH P gingivalis rFimA GAPDH BIAcore K 10 10 M 779 S oralis GAPDH rGAPDH rFimA 434X10 M 6 75 X10 MT P gingivalis
13. kass M s ka s KM S oralis GAPDH rFimA 2 65 x 104 6 11 x 10 4 34 x 107 S oralis TGAPDH rFimA 7 41 x 10 1 10 x 10 6 75 x 107 3 P gingivalis GAPD HH S oralis ATCC 9811 S oralis ATCC 10557 S gordonii GOB S sanguinis ATCC 10556 S parasanguinis ATCC 15909 S sobrinus B 13 S sobrinus 6715 S salivarius OMZ 65 S salivarius HHT S mutans MT 8148 S cricetus HS 6 S milleri NCTC 10703 S sobrinus K1 9 S cricetus HS 1 o n 3 P O e e e e e N N WW WD Q GAPDH 10 M NADH min TIEI 3 9 0 6 5 8 0 9 4 1 0 4 5 8 0 9 2 1 2 12 3 2 0 5 2 6 0 1 33d 3513 3442 1 3 1 04 2 6 0 4 1 3 0 6 0 4 0 2 100 0 0 1 2 3 4 5 ug ml 3 S oralis ATCC 9811 pH GAPDH rGAPDH P gingivalis 381 S oralis ATCC 9811 n 3 RIS pH GAPDH O rGAPDH S pneumoniae 1 MICSDSSYSFHNKNFMIFIRRKSLMVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLT 60 pyogenes rsen deed MVVKVGINGFGRIGRLAFRRIQNIEGVEVTRINDLT 36 gordoni ae ee KVGINGFGRIGRLAFRR
14. SspA SspB 3 A naeslundii 200kDa AnAF P gingivalis P gingivalis c OOCRESEAZ IA l BST 2 HE ETE 1 P gingivalis 381 S oralis ATCC 9811 P gingivalis 381 gingivalis 381 S oralis ATCC 9811 2 L 3 P gingivalis P gingivalis 381 S oralis ATCC 9811 LAL
15. S oralis ATCC 9811 GAPDH DNA M13 20 5 GTAAAACGACGGCCAG 3 M13 5 CAGGAAACA GCTATGAC 3 Sof2 5 GTAGTTAAAGTTGGTATT 3 Sof3 5 CGTTTCGACGGTACT 3 Sor2 5 ACCGTC AACGTCAAGAA 3 Sor3 5 GCAGCACCAGTTGA 3 OD 4 SIGMA Genosys Japan DYEnamic ET Terminator Cycle Sequencing Kit Amersham Biosciences 0 2 e pmol 8 EE 20 u 9 1 9 20 50C 15 60C 60 25 SigmaSpin Postreaction Purification Sigma 4 ABI Prism 310 Genetic Analyzer PE Biosystems go DIG High Prime DNA Labeling and Detection Starter Kit II Roche Diagnostics Mannheim Germany
16. GAPDH 14 G 3 P 40 mM 50 mM 5 mM EDTA pH 8 6 NAD NAD NADH Ko 5 GAPDH CBB GAPDH SDS PAGE rFimA rFimA rFimA A CBB B 26 Veal 2 S oralis ATCC 9811 3 S oralis ATCC 10557 4 S gordonii GIB 5 S sanguinis ATCC 10556 6 S parasanguinis ATCC 15909 10 S oralis ATCC 9811 GAPDH 4 GAPDH 5 5 3 4
17. TERME SERA TOKE ZA S oralis ATCC 9811 rGAPDH P gingivalis 381 S oralis ATCC 10557 S gordonii GOB S sanguinis ATCC 10556 S parasanguinis ATCC 15909 D GAPDH p gingivalis DHEK L LT P gingivalis A naeslundii KWS 81 AnAF S gordonii GOB SspB Yamaguchi It A naeslundii KWS 81 AnAF A naeslundii KWS 81 P gingivalis KC 409 IREE e DHR ZIMEL AnAF Lk AnAF P gingivalis 381 oralis ATCC 98
18. P gingivalis P gingivalis C P gingivalis Actinomyces naeslundii 9 Actinomyces viscosus 1532 33 40 Streptococcus gordonii Streptococcus mutans Streptococcus gs Streptococcus sanguis 7 P gingivalis A viscosus S gordoni L DIHET A Z L IEZ A naeslundii A viscosus S gs 4 viscosus P gingivalis S gordonii antigen I II family
19. Nakamura P gingivalis K 2 15X10 M 216X10 M a 226x106 M I 2 76X10 M 3 08 X 10 MT 3 79X10 M Amano P 15 gingivalis 1 62 X 10 M 1 48 X 10 M7 2 16x 105 M 2 43 K S oralis GAPDH amp P gingivalis GAPDH PE 1 63 x 10 M 10M 14
20. 8 M 50 mM pH 8 0 Sepharose CL 6B 2 5X150 em Pharmacia L rGAPDH Lae 12 S oralis GAPDH P gingivalis P gingivalis rFimA S oralis ATCC 9811 GAPDH BIAcore2000 BIAcore Uppsala Sweden CMS BIAcore M_ WM A 3 u min 37C 10 mM MERT FU v AXE pH 4 8 S oralis ATCC 9811 GAPDH rGAPDH 20 ug m 650X 40kDal 41kDa RU 1 M 10 mM
21. C S oralis ATCC 9811 S oralis ATCC 9811 COMED OPEL VU RAO RIA P gingivalis 1 P gingivalis 381 Forma Scientific Anaerobic System Model 1024 Marietta OH U S A 10 CO 10 Ho 80 N gt 35 2 1 mg ml BBL
22. S oralis GAPDH PCR pBT3d Novagen Inc Madison Wis U S A NcoI BamHI L 5 oralis GAPDH LEE coli BL21 DE3 LB 1 mM D C IPTG rGAPDH 2 500Xg 15 50 mM pH 8 0 10 000Xg 30 1 M PE 2 Triton X 100 10 mM EDTA 50 mM F Y A RETE pH 8 0 12 RIAL Z 10 000 X g 30 8 M 0 1 mM PMSF 50 mM pH 8 0 13 500Xg 30
23. S oralis GAPDH GAPDH REA IE GAPDH GAPDH S oralis GAPDH GAPDH G 3 P 1 3 4 EROR Pancholi D lx Z jv 7 A GAPDH A GAPDH
24. 16 DNA S oralis ATCC 9811 S oralis ATCC 10537 S gordonii GOBER S sanguinis ATCC 10556 5S parasanguinis ATCC 15909 GAPDH DDBJ AB110908 AB163424 AB163425 AB163426 AB163427 S pneumoniae TIGR4 7 S gordonii FSS2 40 S pyogenes M1 S equisimilis H46A K Staphylococcus epidermidis ATCC 122288 Staphylococcus aureus N315 10 k QO Fusobacterium nucleatum ATCC 25586 AE007490 AF247678 AE006494 X97788 AE016745 AP003360 AEBO10576 c H Ao X 1 S oralis ATCC 9811 RO FERE SR O HF XE P gingivalis FimA S oralis ATCC 9811 S oralis ATCC 9811 ROHE IL 40kDa 1 4 2 S oralis ATCC 9811 S oralis ATCC 9811 RO H PERSE Z EE KALE CHAPS CNBr activated Sepharose 4B
25. Sof2 Sor2 25 ng ml PCR 909 bp DNA DIG S oralis ATCC 9811 DNA 4 ug GAPDH 10 BamHI EcoRI HindIII Kpnl Pstl Sacl Sall Smal Sphl Xbal New England Biolabs 37C 20 7 XSSC 1XSSC 0 05 M 0 1S M Roche Diagnostics DNA UV 10 ml DIG Easy Hyb 42 30 DIG 87 5 ng 3 5 ml DIG Easy Hyb 42 0 1 SDS 2X SSC 2
26. 10 mM PBS 25 000Xg 30 30 4 3 25 000 X g 30 2 M 50 mM pH 7 5 IC CHES EH LER P gingivalis rFimA CNBr activated Sepharose 4B Amersham Pharmacia Biotech AB Uppsala Sweden 4 rFimA Amersham Pharmacia Biotech 0 5 M rFimA rFimA S oralis ATCC 9811 BERRE L SDS PAGE Y F 5 5 ug SDS PAGE 1 PBS
27. Sofl Sofl PCR 14 T J 1 1kbp PCR Z GAPDH HARS GAPDH ME amp FERRE IEE t OTIC AR ZF 14 GAPDH GAPDH GAPDH GAPDH P gingivalis GAPDH js b JCGEEEISTE2 CHRV DES V LOVER A GAPDH S oralis ATCC 9811 GAPDH P gingivalis 5
28. Streptococcus sobrinus K1 9 B 13 fk 6715 Streptococcus salivarius OMZ 65 HHT Streptococcus cricetus HS 1 HS 6 S oralis ATCC 9811 448 AIL 5 000 X g 30 0 15 M 10 mM PBS phosphate buffered saline pH 6 0 3 yee yey UT Escherichia coli JM109 W BL21 DE3 Novagen Madison WI U S A Luria Bertani LB 1 mg ml 1 mg ml 0 5 mg ml LB 50 ng ml 2 SDS SDS PAGE sodium dodecyl sulfate poly acrylamide gel electrophoresis Laemmli 0 1 SDS 12 5 2 SDS 6 8
29. rFimA 1 mg ml 4C 0 05 Tween 20 PBS 3 400 rFimA 4 2 000 HRP IsG H L Zymed Laboratories Inc San Francisco CA U S A 1 HRP Conjugate Substrate Kit Bio Rad rEimA Lae Prestrained SDS PAGE Standards Low Range Bio Rad B 112 kDa BSA 81 kDa 49 9 kDa kx 36 2 kDa 29 9 kDa 21 3 kDa 6 Nagata Cisar 10 mM PBS pH 6 0 2 ml 5x105 P gingivalis 381 Sx10 S oralis ATCC 9811 UV 265FW 37 C CHEE L728 6 550nm 4s 7 5
30. 1 2 orali 3 S oralis 4 rFimA S oralis 2 S oralis ATCC 9811 PRO FE SUERTE GS pH GAPDH rGAPDH CBB 5 ug amp SDS PAGE 12 5 rFimA 1 mg ml rFimA lsG HRP conjugate lsG rFimA HRP Conjugate Substrate Kit A CBB B 1 3 pH GAPDH 4 rGAPDH P gingivalis 381 S oralis ATCC 9811 2 3 S oralis ATCC 9811 FRO FRE SEER SR pH GAPDH rGAPDH C X 10 mM PBS pH 6 0 2 ml 5x10 D P gingivalis 381 Sx10 S oralis ATCC 9811
31. PCR 6 S oralis ATCC 9811 Puregene DNA Isolation Kit Gentra Systems Minneapolis Minn U S A DNA 10 ng 50 mM 10 mM bY A EET ET ik pH 8 3 2 5 mM 200 uM 3 1 0 uM Sofl Sorl EU 0 025 U AmpliTaq Gold DNA zh 100 ul amp 94 9 94 45 50 1 72 8 Ine 33 PCR 1 143 bp pCR2 1 TOPO TA Cloning Kit Invitrogen Carlsbad CA U S A E coi TOPO10 X gal mvitrosgen LB Sofl Sor1 PCR EcoRI New England Biolabs Beverly Mass U S A
32. 1 P gingivalis 381 Li A B A x100 A B 5x10 P gingivalis 381 5x105 1 mM 0 1 mM 0 15 M 0 1 mM 0 02 pH 7 2 10 0 1 2 RARR TEE CH 523 5 3 4 5 7 S oralis ATCC 9811 SDS PAGE PVDP Tra
33. 62 5 mM pH 6 8 30 SDS k Bio Rad Bio Rad Laboratories Hercules CA U S A 25 mA LS CBB Coomassie Brilliant Blue SDS LMW Calibration Amersham Biosciences Piscataway N J U S A B 97 kDa 66 kDa 45 kDa 30 kDa 20 1 kDa oc 14 4 kDa BCA Protein Assay Kit Pierce Rockford Ill U S A BSA bovine serum albumin 3 S oralis ATCC 9811 Nagata P gingivalis rFimA 3 S oralis Lamont
34. mM NADH min 14 GAPDH GAPDH S oralis ATCC 9811 S oralis ATCC 10557 S gordonii G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 5 GAPDH Winram 30 1 mM PMSF 1 mM AM L TLCK Sigma PBS 180 U g 37C 2 25 000Xg 30 30 kDa MW cut off Amicon Cibacron Blue Sepharose CL 6B KARO GAPDH 15 GAPDH SPSS ver 10
35. 0 1 g 4 g 1g 10 g L 0 5 g 2N pH 7 5 6 000 Xs 20 50 mM bY AERA EHE pH 7 3 CHT L 30 kDa MW cut off Amicon Millipore Corp Bedford Mass U S A Cibacron Blue Sepharose 3 0X15 cm Amersham Pharmacia Biotech GAPDH NAD 11 S oralis GAPDH GAPDH GAPDH Kuboniwa Acol GCCCATGG 8 TAGTTAAAGTTGGTATTAACGGT 3 PamHI 85 GCGGATCCTTATTTAGCGATTTTTGCGAAGT 3 S oralis GAPDH PCR
36. Role of Porphyromonas gingivalis 40 kDa outer membrane protein in the aggregation of P gingivalis vesicles and Actinomyces viscosus Arch Oral Biol 37 717 724 18 Holland J P 1983 Homologous nucleotide sequence at the 5 termini of messenger RNAs synthesized from the yeast enolase and glyceraldehyde 3 phosphate dehydrogenase gene families J Biol Chem 258 5291 5299 19 Isogai H Isogai E Yoshimura F Suzuki T Kagota W and Takano K 1988 Specific inhibition of adherence of an oral strain of Bacteroides gingivalis 381 to epithelial cells by monoclonal antibodies against the bacterial fimbriae Arch Oral Biol 33 479 485 20 Jenkinson H F and Demuth D R 1997 Structure function and immunogenicity of streptococcal antigen I II polypeptides Mol Microbiol 23 183 190 21 Kamaguchi A Baba H Hoshi M and Inomata K 1995 Effect of Porphyromonas 20 gingivalis ATCC 33277 vesicle on adherence of Streptococcus mutans OMZ 70 to the experimental pellicle Microbiol Immunol 39 521 524 22 Kapatral V Anderson I Ivanova N Reznik G Los T Lykidis A Bhattacharyya A Bartman A Gardner W Grechkin G Zhu I Vasieva O Chu L Kogan Y Chaga O Goltsman E Bernal A Larsen N D Souza M Walunas T Pusch G Haselkorn R Fonstein M Kyrpides N and Overbeek R 2002 Genome sequence and analysis of the oral bacterium Fusobacterium nucleatum
37. oralis ATCC 9811 RO FERRE EEE SR IL 40kDa th S pneumoniae S gordonii S pyogenes S equisimilis GAPDH 90 S aureus S epidermidis GAPDH 70 GAPDH 17 E nucleatum 3 S oralis ATCC 9811 GAPDH rGAPDH P gingivalis BIAcore GAPDH K 2434 X 10 M rGAPDH 6 75X107M D 4 14 GAPDH r 0 854 P lt 0 01 1 5 GAPDH pE amp IEE LE AIELLO ERE 4 GAPDH 40kDa GAPDH rEimA GAPDH S oralis AT
38. Osaka University Knowledg Streptococcus oralig glyceral dehyde 3 phosphate Title dehydrogenasd Porphyromonas gingivalis IlHHHHHHHLH Author s Citation Issue Date Text Version ETD URL http hdl handlene 11094 880 DOI Rights Osaka University Knowledge Archive OUKA http ir library osaka u ac jp dspace Osaka University Streptococcus oralis glyceraldehyde 3 phosphate dehydrogenase 3 Porphyromonas gingivalis E Tul Porphyromonas gingivalis 080999 ASTA IA E OPE ET OT Hi P gingivalis WE
39. B 0 1 SDS 0 5XSSO 68C 2 C 150 mM 0 3 Tween20 100 mM 5 1 150 mM 100 mM 1 DIG 150 mU mD 30 C 15 0 1 M pH 9 5 0 1 M CDP Star disodium 2 chloro 5 4 methoxyspiro 1 2 dioxetane 3 2 5 chloro tricycle 3 3 1 1 decan 4 yl 1 phenyl phosphate 10 S oralis ATCC 9811 pH GAPDH Nelson pH S gordonii GAPDH S oralis ATCC 9811 GAPDH S oralis ATCC 9811 11 22 5 g 2 9
40. Becton Dickinson Microbiology Systems Cockeysville MD U S A 5 ug ml Sigma Sigma Aldrich Japan K K R 1 pg ml Sigma 30 g l BBL 10 ml 35C 24 S oralis ATCC 9811 American Type Culture Collection Rockville MD U S A BHI brain heart infusion broth BBL 10 ml 35 C 15 WRF Ae Lic 100 ml 3 000 ml S oralis ATCC 10557 S sanguinis ATCC 10556 Streptococcus parasanguinis 2 ATCC 15909 American Type Culture Collection S gordonii GOB Genco mutans MT 8148 Streptococcus milleri 10703 National Collection of Type Cultures London England
41. 0 60 N Procise cLC PE Biosystems Branchburg N J U S A 8 GAPDH S oralis ATCC 9811 N DDBJ http srs ddbj nig ac jp FASTA Streptococcus pyogenes Streptococcus equisimilis Streptococcus pneumoniae S gordonii glyceraldehyde 3 phosphate dehydrogenase GAPDH 4 AEO GAPDH Sofl 5 AGTTCTGTTGAAAGG 3 Sorl 5 CGAAAAAGAACTCAGC 3 SIGMA Genosys Japan S oralis ATCC 9811 FROOJCEEE SR 3E SY n vZ S oralis PCR Gene Amp 2400 PE Biosystems
42. 300 300 MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV xoleloleelolololelelelelelololeleleleelotoleleleleletotoleleteleletetotoleleleleetotoleleleleetotoleleleleletotolol KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKAAAEKHLHAGGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I KEGGFEVNGKF VKVSAERDPENTDWANDGVE I VLEATGF F ATKAAAEKHLHAGGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I xoleloleelolololelelelelelololelelelelek lololok xololelelelelelelololelelek ok kk xlolelelelololok TAPGGNDVKTVVFNTNHDVLDGTETV ISGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV I SGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGSDVKTVVFNTNHDILDGTETV I SGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV I SGASCT TNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV ISGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA xolololek xololelelelelelololelek oelololelelelelelotoleleleleleteloteleteleleetotoleleleleetotoleleleleletotoloe YTGDQMILDGPHRGGDLRRARAGAAN I VPNSTGAAKA T GLV I PELNGKLDGSAQRVPTPT YTGDQMILDGPHRGGDLRRARAGAVN I VPNSTGAAKA I GLV I PELNGKLDGAAQRVPVPA YTGDQMILDGPHRKGDLRRARAGAAN I VPNST
43. A major surface protein on group A streptococci is a glyceraldehyde 3 phosphate dehydrogenase with multiple binding activity J Exp Med 176 415 426 45 Pancholi V and Fischetti V A 1993 Glyceraldehyde 3 phosphate dehydrogenase on the surface of group A streptococci is also an ADP ribosylating enzyme Proc Natl Acad Sci USA 90 8154 8158 46 Rickard A H Gilbert P High N J Kolenbrander P E and Handley P S 2003 Bacterial coaggregation an integral process in the development of multi species biofilms Trends Microbiol 11 94 100 47 Schwarz S Ellen R P and Grove D A 1987 Bacteroides gingivalis Actinomyces viscosus cohesive interactions as measured by a quantitative binding assay Infect Immun 55 2391 2397 48 Slots J and Genco R J 1984 Black pigmented Bacteroides species Capnocytophaga species and Actinobacillus actinomycetemcomitans in human periodontal disease Virulence factors in colonization survival and tissue destruction J Dent Res 63 412 421 49 Slots J and Gibbons R J 1978 Attachment of Bacteroides melaninogenicus subsp asaccharolyticus to oral surfaces and its possible role in colonization of the mouth and of periodontal pockets Infect Immun 19 254 264 50 Stinson M W Safulko K and Levine M J 1991 Adherence of Porphyromonas Bacteroides gingivalis to Streptococcus sanguis in vitro Infect Immun 59 102 108 51 Tettelin H
44. ATCC 10557 S gordonii GOBER t f 1 0 j l 5 0 1 2 3 4 5 0 1 3 A S oralis rGAPDHig ug ml S oralis rGAPDHig ug ml S oralis rGAPDH pg ml S parasanguinis ATCC 15909 S sanguinis ATCC 10556 1 1 i 1 S o7g7 srGAPDH ug ml S oralis rGAPDH ug ml 11 S oralis ATCC 9811 rGAPDH P gingivalis 381 n 3 1 S oralis ATCC 9811 S oralis ATCC 9811 WV 3 mg 10 rEimA gsG 1 500 Z ASAN L Affi Gel A ug SDS PAGE P gingivalis rFimA S oralis ATCC 9811 150 ng rFimA 50 ng 10 HRP Conjugate HRP Conjugate Substrate Kit
45. Nelson K E Paulsen I T Eisen J A Read T D Peterson S Heidelberg J DeBoy R T Haft D H Dodson R J Durkin A S Gwinn M Kolonay J F Nelson W C Peterson J D Umayam L A White O Salzberg S L Lewis M R Radune D Holtzapple E Khouri H Wolf A M Utterback T R Hansen C L McDonald L A Feldblyum T V Angiuoli S Dickinson T Hickey E K Holt L E Loftus B J Yang F Smith H O Venter J C Dougherty B A Morrison D A Hollingshead S K and Fraser C M 2001 Complete genome sequence of a virulent isolate of Streptococcus pneumoniae Science 293 498 506 52 van Winkelhoff A J van der Velden U and de Graaff J 1987 Microbial succession in recolonizing deep periodontal pockets after a single course of supra and subgingival debridement J Clin Periodontol 15 116 122 23 53 Winram S B and Lottenberg R 1996 The plasmin binding protein Plr of group A streptococci is identified as glyceraldehyde 3 phosphate dehydrogenase Microbiology 142 2311 2320 54 Yamaguchi T Kasamo K Chuman M Machigashira M Inoue M and Sueda T 1998 Preparation and characterization of an Actinomyces naeslundii aggregation factor that mediates coaggregation with Porphyromonas gingivalis J Periodontal Res 33 460 468 24 kDa 112 81 49 9 40 kDa 36 2 29 9 21 3 1 eS
46. oralis ATCC 9811 i oralis HARE RS HH 3 oralis 4 rFimA oralis kDa kDa 97 112 66 81 45 r 49 9 30 36 2 29 9 20 1 14 4 21 3 1 2 3 4 1 2 2 S oralis ATCC 9811 ORN H ES S pH CBB A CBB B 1 2 3 pH GAPDH 4 rGAPDH 3 4 GAPDH rGAPDH 1 S oralis GAPDH Streptococcus oralis Streptococcus pheumoniae Streptococcus oralis Streptococcus parasanguinis Streptococcus sanguinis Streptococcus gordonii Streptococcus gordonii Streptococcus pyogenes Streptococcus equisimilis Staphylococcus epidermidis Staphylococcus aureus Fusobacterium nucleatum ATCC 9811 TIGR4 ATCC 10557 ATCC15909 ATCC 10556 FSS2 G9B MI GAS H46A ATCC 12228 N315 ATCC 25586 100 977 96 95 95 93 93 89 88 71 71 66 100 99 98 98 97 97 97 92 88 70 70 71 R2 BlAcorel 4 amp 6S oralis GAPDHEP gingivalis
47. 10 M NADH min 3 9X S cricetus HS 1 S sobrinus K1 9 GAPDH 1 3 10 M NADH min HRE L ARES GAPDH P lt 0 01 GAPDH P gingivalis amp 9R lt Hist GAPDH Bd 8 r 0 854 13 12 GAPDH GAPDH S oralis ATCC 9811 S oralis ATCC 10557 f S gordonii G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 GAPDH SDS PAGE 40kDa GAPDH p gingivalis rFimA Ed 9 13 GAPDH S oralis ATCC 10
48. 11 Lamont S gordonii GOB 170kDa SspB P gingivalis 33277 16 SspB 1167 1193 P gingivalis 2 Chung LS gordonii SspB P gingivalis 100kDa P gingivalis P gingivalis S gordonii SspB P gingivalis S oralis SEER amp AE L GAPDH S ozg GAPDH p gingivalis
49. 557 S gordonii G9B S sanguinis ATCC 10556 parasanguinis ATCC 15909 GAPDH 5 GAPDH 10 5 GAPDH oralis ATCC 9811 GAPDH S oralis ATCC 10557 S gordonii G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 GAPDH 97 6 97 0 97 0 97 6 1 SS 14 S oralis ATCC 9811 rGAPDH P gingivalis 381 5 GAPDH P gingivalis P gingivalis 381 5 S oralis ATCC 9811 rGAPDH S oralis ATCC 9811 rGAPDH T 2 ug ml Bd 11
50. AGAVN I VPNSTGAAKA I GRV I PELNGKLDGAAQRVPVPA YTGDQMILDGPHRGGDLRRARAGAVN I VPNSTGAAKA I GRV I PELNGKLDGAAQRVPVPA Joleloloelolololelolelek xolelolelelelololek xelelololelelelelelololelek xelolololelelelelolok xlekelelok k GSVTELVAVLEKNVTVDEVNAAMKAASNESYGYTEDP I VSSDIVGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDP I VSSDILGMSYGSLFDATQTKVL GSVTELVVVLEKNVTVDEVNAAMKAVANESYGYTEDP I VSSDVVGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDPIVSSDILGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDPIVSSDILGMSYGSLFDATQTKVL xolelolelelek k xololelelelelelololelek ololelelelelelelololelelelek xelolelelelololelelelelelololol DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK 60 60 60 60 60 120 119 120 119 119 180 179 180 179 179 240 239 240 239 239 300 299 300 299 299 336 335 336 335 335 10 S oralis ATCC 9811 GAPDH 4 GAPDH 3 4 S oralis ATCC 9811 S oralis
51. CC 9811 GAPDH 97 6 S oralis ATCC 9811 rGAPDH P gingivalis 381 4 2 ug ml P gingivalis S oralis ATCC 9811 7 JETER 40kDa GAPDH CHS 7 E 238 572 E 72 0 P gingivalis P gingivalis GAPDH
52. DH P gingivalis 381 kDa A kDa B 112 97 WW g1 66 mm 45 Luo 49 9 30 a 36 2 20 1 29 9 14 4 a 21 3 1 2 3 45 6 12 3 4 5 6 9 s GAPDH CBB A CBB B 1 2 8 oralis ATCC 9811 3 S oralis ATCC 10557 4 S gordonii G9B 5 S sanguinis ATCC 10556 6 S parasanguinis ATCC 159097 S ora s ATCC 9811 oralis ATCC 10557 S gordoni i G9B sanguinis ATCC 10556 parasanguinis ATCC 15909 S oralis ATCC 9811 S oralis ATCC 10557 S gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 oralis ATCC 9811 S oralis ATCC 10557 gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 S oralis ATCC 9811 oralis ATCC 10557 S gordoni i G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 ora s ATCC 9811 oralis ATCC 10557 S gordoni G9B sanguinis ATCC 10556 S parasanguinis ATCC 15909 S oralis ATCC 9811 oralis ATCC 10557 S gordoni G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 61 61 61 61 61 121 120 121 120 120 181 180 181 180 180 241 240 241 240 240 301 300 301 300 300 MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLA
53. GAAKA T GLV I PELNGKLDGAAQRVPTPT YTGDQMILDGPHRGGDLRRARAGAVN I VPNSTGAAKA I GRV I PELNGKLDGAAQRVPVPA YTGDQMILDGPHRGGDLRRARAGAVN I VPNSTGAAKA I GRV I PELNGKLDGAAQRVPVPA Joleloloelolololelolelek xolelolelelelololek xelelololelelelelelololelek xelolololelelelelolok xlekelelok k GSVTELVAVLEKNVTVDEVNAAMKAASNESYGYTEDP I VSSDIVGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDP I VSSDILGMSYGSLFDATQTKVL GSVTELVVVLEKNVTVDEVNAAMKAVANESYGYTEDP I VSSDVVGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDPIVSSDILGMSYGSLFDATQTKVL GSVTELVAVLEKSVTVDEVNAAMKAASNESYGYTEDPIVSSDILGMSYGSLFDATQTKVL xolelolelelek k xololelelelelelololelek ololelelelelelelololelelelek xelolelelelololelelelelelololol DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK DVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAK I AK 60 60 60 60 60 120 119 120 119 119 180 179 180 179 179 240 239 240 239 239 300 299 300 299 299 336 335 336 335 335 10 S oralis ATCC 9811 GAPDH 4 GAPDH 3 4
54. HLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEV xoleloleelolololelelelelelololeleleleelotoleleleleletotoleleteleletetotoleleleleetotoleleleleetotoleleleleletotolol KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKAAAEKHLHAGGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I KEGGFEVNGKF VKVSAERDPENTDWANDGVE I VLEATGF F ATKAAAEKHLHAGGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I KEGGFEVNGKF VKVSAERDPEQTDWATDGVE I VLEATGF F AKKDAAEKHLK GGAKKVV I xoleloleelolololelelelelelololelelelelek lololok xololelelelelelelololelelek ok kk xlolelelelololok TAPGGNDVKTVVFNTNHDVLDGTETV ISGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV I SGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGSDVKTVVFNTNHDILDGTETV I SGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV I SGASCT TNCLAPMAKALQDNF GVVEGLMTT IHA TAPGGNDVKTVVFNTNHDVLDGTETV ISGASCTTNCLAPMAKALQDNF GVVEGLMTT IHA xolololek xololelelelelelololelek oelololelelelelelotoleleleleleteloteleteleleetotoleleleleetotoleleleleletotoloe YTGDQMILDGPHRGGDLRRARAGAAN I VPNSTGAAKA T GLV I PELNGKLDGSAQRVPTPT YTGDQMILDGPHRGGDLRRARAGAVN I VPNSTGAAKA I GLV I PELNGKLDGAAQRVPVPA YTGDQMILDGPHRKGDLRRARAGAAN I VPNSTGAAKA T GLV I PELNGKLDGAAQRVPTPT YTGDQMILDGPHRGGDLRRAR
55. Hllen R P and Grove D A 1989 Bacteroides gingivalis vesicles bind to and aggregate Actinomyces viscosus Infect Immun 57 1618 1620 12 Ellen R P Song M and Buivids L A 1992 Inhibition of Actinomyces viscosus Porphyromonas gingivalis coadhesion by trypsin and other proteins Oral Microbiol Immunol 7 198 203 13 Figge R M Schubert M Brinkmann H and Cerff R 1999 Glyceraldehyde 3 phosphate dehydrogenase gene diversity in eubacteria and eukaryotes Evidence for intra and inter kingdom gene transfer Mol Biol Evol 16 429 440 14 Gase K Gase A Schirmer H and Malke H 1996 Cloning sequencing and functional overexpression of the Streptococcus equisimilis H46A gapC gene encoding a glyceraldehyde 3 phosphate dehydrogenase that also functions as a plasmin ogen binding protein Purification and biochemical characterization of the protein Eur J Biochem 239 42 51 15 Goulbourne P A and Ellen R P 1991 Evidence that Porphyromonas Bacteroides gingivalis fimbriae function in adhesion to Actinomyces viscosus J Bacteriol 173 5266 5274 16 Gozalbo D Gil Navarro I Azorin I Renau Piqueras I Martinez J P and Gil M L 1998 The cell wall associated glyceraldehyde 3 phosphate dehydrogenase of Candida albicans is also a fibronectin and laminin binding protein Infect Immun 66 2052 2059 17 Hiratsuka K Abiko Y Ito T Sasahara H and Takiguchi H 1992
56. IQNVEGVEVTRINDLT 33 S QUIETIS MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLT 36 oralis a See ee MVVKVGINGFGRIGRLAFRRIQNVEGVEVTRINDLT 36 S pneumoniae 61 DPVMLAHLLKYDTTQGRFDGTVEVKEGGFEVNGKFIKVSAERDPEQIDWATDGVEIVLEA 120 pyogenes 37 DPNMLAHLLKYDTTQGRFDGTVEVKEGGFEVNGNF IKVSAERDPEN I DWATDGVE I VLEA 96 S gordoni 34 DPVMLAHLLKYDTTQGRFDGTVEVKEGGFEVNGKFVKVSAERDPEN I DWANDGVE I VLEA 93 S equ isimilis 31 DPNMLAHLLKYDTTQGRFDGTVEVKEGGFEVNGNF IKVSAERDPENIDWATDGVE I VLEA 96 oralis 37 DPVMLAHLLKYDTTQGRFDGTVEVKEGGFEVNGKF VKVSAERDPEQ I DWATDGVE I VLEA 96 xk solelelelelelotelolelelelelelotelelelelelelelelelelelelelek ok xolelelelelelelek xlelelek slolelelelelelelek S pneumoniae 121 TGFFAKKEAAEKHLK GGAKKVVITAPGGNDVKTVVFNTNHDVLDGTETVISGASCTTNC 179 pyogenes 97 TGFFAKKEAAEKHLHANGAKKVV ITAPGGNDVKTVVFNTNHDILDGTETVISGASCTTNC 156 S gordoni i 94 TGFFATKAAAEKHLHAGGAKKVVITAPGGSDVKTVVFNTNHDVLDGTETVISGASCTTNC 153 S equisimi is 91 TGFFAKKEAAEKPLHANGAKKVVITAPGGNDVKQLFSTLTTSILDGTETVISGASCTTNC 156 S oralis 97 TGFFAKKAAAEKHLHAGGAKKVV I TAPGGNDVKTVVFNTNHDVLDGTETVISGASCTTNC 156 xoblelek k kkk k KKK S pneumoniae 180 LAPMAKALQDNFGVVEGLMTT IHAYTGDQMILDG
57. PDH 1 10 X 10 s E D 675x10 M K GAPDH 4 34 X 10 M rGAPDH 10 P gingivalis 14 12 P gingivalis 381 WR amp Hee Lie 3 S oralis ATCC 9811 P gingivalis 381 S oralis ATCC 10557 S gordonii G9B S sanguinis NTCC 10556 S parasanguinis ATCC 15909 P gingivalis 381 Fk amp DROED A BATE S sobrinus K1 9 S cricetus HS 1 ll GAPDH GAPDH S oralis GAPDH AAO ATCC 9811 GAPDH 7 7X10 M NADH min 3 S oralis ATCC 10557 S gordonii G9B S sanguinis ATCC 10556 S parasanguinis ATCC 15909 GAPDH 1f
58. PHRGGDLRRARAGAANIVPNSTGAAK 239 pyogenes 157 LAPMAKALHDAFGIQKGLMTTIHAYTGDQMILDGPHRGGDLRRARAGAANIVPNSTGAAK 216 S gordoni 154 LAPMAKALQDNFGVVEGLMTTIHAYTGDQMVLDGPHRKGDLRRARAAAANIVPNSTGAAK 213 equisimi is 151 LAPMAKALHDAFGIQKGLMTTIHAYTGDQMIVDGHRGGGDLRRARAGAANIVPNSTGARK 216 oralis 157 LAPMAKALQDNFGVVEGLMTT IHAYTGDQMILDGPHRGGDLRRARAGAANIVPNSTGAAK 216 ek ek kkk k S pneumoniae 240 AIGLVIPELNGKLDGSAQRVPTPTGSVTELVAVLEKNVTVDEVNAAMKAASNESYGYTED 299 pyogenes 217 AIGLVIPELNGKLDGAAQRVPVPTGSVTELVVTLDKNVSVDEINAAMKAASNDSFGYTED 276 S gordoni 214 AIGLVIPELNGKLDGAAQRVPVPTGSVTELVAVLDKNVTVDEVNAAMKAASNESYGYTED 273 S equisimi is 211 AIGLVIPELNGKLDGAAQRVPVPTGSVTELVVTLDKNVSVDEINAAMKAASNDSFGYTED 276 S oralis 217 AIGLVIPELNGKLDGSAQRVPTPTGSVTELVAVLEKNVTVDEVNAAMKAASNESYGYTED 276 xololelolelelelelelolololelek kk xlelelelolelolelek ak leek k xlolelelelelelelek ok kkk S pneumoniae 300 PIVSSDIVGMSYGSLFDATQTKVLDVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAKIAK 359 pyogenes 277 PIVSSDIVGVSYGSLFDATQTKVMEVDGSQLVKVVSWYDNEMSYTAQLVRTLEYFAKIAK 336 gordoni 214 PIVSSDIVGMAYGSLFDATQTKVLDVDGKQLVKVVSWYDNEMSYTAQ 320 S equisimilis 211 PIVSSDIVGVSYGSLFDATQTKVMEVDGSQLVKVVSWYDNEMSYTAQLVRTLEYFAKTAK 336 S oralis 211 PIVSSDIVGMSYGSLFDATQTKVLDVDGKQLVKVVSWYDNEMSYTAQLVRTLEYFAKIAK 336 ololeleletelelek oloteleletelelelelele
59. S oralis ATCC 9811 5 mM 2 mM 1 mM pH 6 0 0 1 M pH B DICRMLTE N t 3 mg 10 Sisma 3 10 000Xg 10 2 REDE GE IRE CRUS L PASTS E UR 200P 200W 1 5 000 X g 30 S oralis 150 ug P gingivalis rFimA 50 ug 2 rFimA IgG 1 500 30 Affi Gel A 10 ug Bio Rad 4 1 Re Le Affi Gel AMAPS II Bio Rad 4 Aff Gel A MAPS II Bio Rad AI SDS PAGE SD Bio Rad
60. eptococcus oralis through specific domains J Dent Res 76 852 857 Amano A 2003 Molecular interaction of Porphyromonas gingivalis with host cells Implication for the microbial pathogenesis of periodontal disease J Periodontol 74 90 96 Arai M Hamada N and Umemoto T 2000 Purification and characterization of a novel secondary fimbrial protein from Porphyromonas gingivalis strain 381 FEMS Microbiol Lett 193 75 81 Brooks W Demuth D R Gil S and Lamont R J 1997 Identification of a Streptococcus gordonii SspB domain that mediates adhesion to Porphyromonas gingivalis Infect Immun 65 3753 3758 Chung W O Demuth D R and Lamont R J 2000 Identification of a Porphyromonas gingivalis receptor for the Streptococcus gordonii SspB protein Infect Immun 68 6758 6762 Cisar J O Kolenbrander P E and McIntire F C 1979 Specificity of coaggregation reactions between human oral streptococci and strains of Actinomyces viscosus or Actinomyces naeslundii Infect Immun 24 742 752 D Costa S S and Boyle M D 2000 Interaction of group A streptococci with human plasmin ogen under physiological conditions Methods 21 165 177 10 Demuth D R Irvine D C Costerton J W Cook G S and Lamont R J 2001 Discrete protein determinant directs the species specific adherence of Porphyromonas gingivalis to oral streptococci Infect Immun 69 5736 5741 19 11
61. ilis GAPDH GAPDH PCR Sofl Sorl 11 PCR 1 1kbp DNA S oralis ATCC 9811 GAPDH 4 1 S oralis GAPDH S pneumoniae S gordonii S pyogenes S equisimilis GAPDH 90 S epidermidis S aureus nucleatum GAPDH 70 5 oralis ATCC 9811 GAPDH GAPDH GAPDH 9 S oralis GAPDH Sof2 Sor2 PCR 909 bp
62. lek oelek olleleleleleteleleleleleleleleleiek 4 S oralis GAPDH GAPDH 5 3 4 N N Kb 15 0 5 0 3 5 1 5 5 BamH I EcoRI Hindi KpnI PstI SacI Sall Smal SphI T T S oralis ATCC 9811 GAPDH Xba I 6 P gingivalis 381 S oralis ATCC 9811 A P gingivalis 381 oralis ATCC 9811TR OH eZ B S oralis rGAPDH 2 5 pg ml Response RU GAPDH rGAPDH 200 100 0 4 uM 150 0 3 uM S ec o L 50 0 2 uM 2 10 o oO 0 1 uM e 50 0 05 uM f 0 4 0 0 50 100 150 200 250 0 7 S oralis GAPDH rFimA GAPDH 105M NADH min 0 1 2 3 4 8 GAP
63. ns blot Transfer Medium 0 2 um Bio Rad CBB N c SDS PAGE CBB 40 10 50 mM pH 8 5 ADC 10 OH 0 1 M bY AYRE RR pH 9 0 10 Speed Vac Savant Instruments Holbrook N Y U S A lug 0 1 M bY Abe ET pH 9 0 0 05 0 Triton X 100 250 ng 37 16 1 TFA 60 4 AX 0 1 TFA Cisa Vydac 218TP5215 2 1 X150 mm 1 ml min 0 1 TFA 60
64. ragments FEMS Microbiol Lett 114 31 36 38 Nagata H Sharma A Sojar H T Amano A Levine M J and Genco R J 1997 Role of the carboxyl terminal region of Porphyromonas gingivalis fimbrillin in binding to salivary proteins Infect Immun 65 422 427 39 Naito Y Tohda H Okuda K and Takazoe I 1993 Adherence and hydrophobicity of invasive and noninvasive strains of Porphyromonas gingivalis Oral Microbiol Immunol 8 195 202 40 Nakamura T Amano A Nakagawa I and Hamada S 1999 Specific interactions between Porphyromonas gingivalis fimbriae and human extracellular matrix proteins FEMS Microbiol Lett 175 267 272 41 Nelson D Goldstein J M Boatright K Harty D W Cook S L Hickman P J Potempa J Travis J and Mayo J A 2001 pH regulated secretion of a glyceraldehyde 3 phosphate dehydrogenase from Streptococcus gordonii FSS2 Purification characterization and cloning of the gene encoding this enzyme J Dent Res 80 371 377 42 Nyvad B and Kilian M 1990 Comparison of the initial streptococcal microflora on dental enamel in caries active and in caries inactive individuals Caries Res 24 267 272 43 Okuda K 1993 Attachment mechanisms and colonization p 139 157 In Shah H N Mayland D and Genco R J ed Biology of the species Porphyromonas gingivalis CRC Press Inc Boca Raton Fla 22 44 Pancholi V and Fischetti V A 1992
65. rsson R E and Rosan B 1993 Involvement of Porphyromonas gingivalis fimbriae in adherence to Streptococcus gordonii Oral Microbiol Immunol 8 272 276 31 Lamont R J Gil S Demuth D R Malamud D and Rosan B 1994 Molecules of Streptococcus gordonii that bind to Porphyromonas gingivalis Microbiology 140 867 872 32 Li J and Ellen R P 1989 Relative adherence of Bacteroides species and strains to Actinomyces viscosus on saliva coated hydroxyapatite J Dent Res 68 1308 1312 21 33 Li J Ellen R P Hoover C I and Felton J R 1991 Association of proteases of Porphyromonas Bacteroides gingivalis with its adhesion to Actinomyces viscosus J Dent Res 70 82 86 34 Mayland D and Holt S C 1988 Biology of asaccharolytic black pigmented Bacteroides species Microbiol Rev 52 134 152 35 Modun B and Williams P 1999 The staphylococcal transferring binding protein is a cell wall glyceraldehyde 3 phosophate dehydrogenase Infect Immun 67 1086 1092 36 Nagata H Murakami Y Inoshita E Shizukuishi S and Tsunemitsu A 1990 Inhibitory effect of human plasma and saliva on co aggregation between Bacteroides gingivalis and Streptococcus mitis J Dent Res 69 1476 1479 37 Nagata H Amano A Hanioka T Tamagawa H Shizukuishi S and Miyata T 1993 Inhibition of coaggregation between Porphyromonas gingivalis and Streptococcus oralis by fibrinogen f
66. s S oralis DHE rGAPDH 2 pg ml GAPDH 3 8 P gingivalis S oralis OOJEEEIEAC 330 4 rGAPDH 12 6A gingivalis 381 S oralis ATCC 9811 FRO FERRER AERTS S oralis rGAPDH 2 5 pg ml P gingivalis 381 S oralis ATCC 9811 IRO HEE i 6B 9 S oralis ATCC 9811 GAPDH P gingivalis rFimA S oralis ATCC 9811 GAPDH P gingivalis rFimA BIAcore rGAPDH rFimA GAPDH 7 kas GAPDH 2 65 X10 Ms rGAPDH 7 41X10 M s ka X GAPDH 6 11 X 10 s rGA
67. strain ATCC 25586 J Bacteriol 184 2005 2018 23 Karlsson R and Falt A 1997 Experimental design for kinetic analysis of protein protein interactions with surface plasmon resonance biosensors J Immunol Methods 200 121 133 24 Khilko S N Jelonek M T Corr M Boyd L F Bothwell A L and Margulies D H 1995 Measuring interactions of MHC class I molecules using surface plasmon resonance J Immunol Method 183 77 94 25 Kolenbrander P E 2000 Oral microbial communities Biofilms interactions and genetic systems Annu Rev Microbiol 54 413 437 26 Kontani M Ono H Shibata H Okamura Y Tanaka T Fujiwara T Kimura S and Hamada S 1996 Cysteine protease of Porphyromonas gingivalis 381 enhances binding of fimbriae to cultured human fibroblasts and matrix proteins Infect Immun 64 756 762 27 Kuboniwa M Amano A and Shizukuishi S 1998 Hemoglobin binding protein purified from Porphyromonas gingivalis is identical to lysine specific cysteine proteinase Lys gingipain Biochem Biophys Res Commun 249 38 43 28 Laemmli U K 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature London 227 680 685 29 Lamont R J Hersey S G and Rosan B 1992 Characterization of the adherence of Porphyromonas gingivalis to oral streptococci Oral Microbiol Immunol 7 193 197 30 Lamont R J Bevan C A Gil S Pe
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