Abstract:To identify the cell wall-binding proteins of Streptococcus suis serotype 2(SS2),according to their structure feature,the substrates of sortase and I type secreted proteins with LysM domain,WxL domain,choline-binding domain,GW domain or S layer homologous domain in the recently published genome of SS2 strain 98HAH33 were firstly identified and then the putative functions were attributed to individual proteins by reference to the identification of conserved domains of InterPro and BlastP servers.Homologous proteins were identified by unfiltered BlastP homology searches(including conserved domain detection).Among the 23 putative proteins with a C-terminal LPXTG recognition signal for covalent attachment to peptidoglycan by sortase,9 with I signal peptide were identified as sortase substrates.Among 9 substrates,YP_001201232,YP_001201531 and YP_001201656 had been experimentally verified to anchor to bacterial cell wall,and YP_001201232 known as the opacity factor of S.suis(OFS)was proved to be the virulence factors.According to function analysis,YP_001201484,YP_001201544,YP_001199825,YP_001197640,YP_001197840 and YP_001199755 appeared to be involved in SS2 pathogenesis.and YP_001200959,YP_001201233 and two proteins with LysM domain(YP_001199784 and YP_001201729)were the hypothetical proteins.These data suggest the majority of putative sortase substrates may implicate in the virulence of SS2 and could serve as a basis for targeted experimental studies into the function of these proteins.
[1]姚火春,陈国强,陆承平.猪链球菌98分离株病原特性鉴定[J].南京农业大学学报,1999,22(2):67-70.
[2]Tang J,Wang C,Feng Y,et al.Streptococcal toxic shock syn-drome caused byStreptococcus suisserotype2[J].PLoS Medi-cine,2006,3(5):151.
[3]Brinster S,Furlan S,Serror P.C-Terminal WxL domain medi-ates cell wall binding inEnterococcus faecalisand other gram-positive bacteria[J].Journal of Bacteriology,2007,189(4):1244-1253.
[4]Marraffini LA,Dedent AC,Schneewind O.Sortases and the art of anchoring proteins to the envelopes of gram-positive bacteria[J].Microbiology and Molecular Biology Reviews,2006,70(1):192-221.
[5]Boekhorst J,de Been MW,Kleerebezem M,et al.Genome-wide detection and analysis of cell wall-bound proteins with LPxTG-like sorting motifs[J].Journal of Bacteriology,2005,187:4928-4934.
[6]Osaki M,Takamatsu D,Shimoji Y,et al.Characterization of Streptococcus suisgenes encoding proteins homologous to sortase of gram-positive bacteria[J].Journal of Bacteriology,2002,184:971-982.
[7]董亚青,王长军,郑峰,等.猪链球菌2型05ZYH33强毒株srt同源序列及srt A原核表达抗原性的分析[J].细胞与分子免疫学杂志,2007,23(5):339-401.
[8]Vanier G,Sekizaki T,Domínguez-Punaro MC,et al.Disruption of srt A gene inStreptococcus suisresults in decreased interac-tions with endothelial cells and extracellular matrix proteins[J].Veterinary Microbiology,2008,127(3-4):417-424.
[9]Wang C,Li M,Feng Y,et al.The involvement of sortase Ain high virulence of STSS-causingStreptococcus suisserotype2[J].Archives of Microbiology,2008.
[10]Vecht U,Wisselink HJ,van Dijk JE,et al.Virulence of Streptococcus suistype2strains in newborn germfree pigs de-pends on phenotype[J].Infection and Immunity,1992,60:550-556.
[11]Baums CG,Kaim U,Fulde M,et al.Identification of a novel virulence determinant with serum opacification activity in Streptococcus suis[J].Infection and Immunity,2006,74:6154-6162.
[12]Li Y,Gottschalk M,Esgleas M,et al.Immunization with re-combinant Sao protein confers protection againstStreptococcus suisinfection[J].Clinical and Vaccine Immunology,2007,14:937943.
[13]Kll L,Krogh A,Sonnhammer EL.Advantages of combined transmembrane topology and signal peptide prediction-the Pho-bius web server[J].Nucleic Acids Research,2007,35(Web Server issue):W429-W432.
[14]Janulczyk R,Iannelli F,Sjohol m AG,et al.Hic,a novel sur-face protein ofStreptococcus pneumoniaethat interferes with complement function[J].The Journal of Biological Chemistry,2000,275(47):37257-37263.
[15]Jarva H,Hell wage J,Jokiranta TS,et al.The group B strep-tococcal beta and pneumococcal Hic proteins are structurally re-latedimmune evasion molecules that bind the complement in-hibitor factor Hin an analogous fashion[J].The Journal of Im-munology,2004,172:3111-3118.
[16]Takamatsu D,Osaki M,Tharavichitkul P,et al.Allelic varia-tion and prevalence of serumopacity factor among theStrepto-coccus suispopulation[J].Journal of Medical Microbiology,2008,57:488-494.
[17]Lindgren PE,McGavin MJ,Signas C,et al.Two different genes coding for fibronectin-binding proteins fromStreptococ-cus dysgalactiae.The complete nucleotide sequences and char-acterization of the binding domains[J].European Journal of Bi-ochemistry,1993,214(3):819-827.
[18]Courtney HS,Hasty DL,Li Y,et al.Serumopacity factor is a major fibronectin-binding protein and a virulence determinant of Mtype2Streptococcus pyogen[J].Molecular Microbiology,1999,32(1):89-98.
[19]Muramatsu H,Tachikui H,Ushida H,et al.Molecular clo-ning and expression of endo--N-acetylglucosaminidase D,which acts on the core structure of complex type asparagine-linked oli-gosaccharides[J].The Journal of Biochemistry,2001,129(6):923-928.
[20]Fontaine MC,Perez-Casal J,Willson PJ.Investigation of a no-vel DNase ofStreptococcus suisserotype2[J].Infection and Immunity,2004,72(2):774-81.
[21]Bethe G,Nau R,Well mer A,et al.The cell wall-associated serine protease Prt A:a highly conserved virulence factor of Streptococcus pneumonia[J].FEMS Microbiology Letters,2001,205(1):99-104.
[22]Itoi Y,Horinaka M,Tsujimoto Y,et al.Characteristic fea-tures in the structure and collagen-binding ability of a ther-mophilic collagenolytic protease fromthe thermophileGeobacil-lus collagenovoransMO-1[J].Journal of Bacteriology,2006,188(18):6572-6579.
[23]Osaki M,Takamatsu D,Shimoji Y,et al.Characterization of Streptococcus suisgenes encoding proteins homologous to sor-tase of gram-positive bacteria[J].Journal of Bacteriology,2002,184(4):971-982.
[24]孙理云.用生物信息学方法预测猪链球菌2型05ZYH33株的脂蛋白[J].微生物学报,2008,48(8):1104-1109.
[25]孙理云,王利巧,毛薇.猪链球菌2型98HAH33株脂蛋白的in silico方法鉴定[J].河南科技大学学报(自然科学版),2008,29:61-65.
[26]Yamamoto S,Muramatsu H,Muramatsu T.Mutational stud-ies on endo-beta-N-Acetyl-glucosaminidase D which hydrolyzes core portion of asparagine-linked complex type oligosaccharides[J].Glycoconjugate Journal,2005,22(1-2):35-42.