Smith Todd G, Lim Jae-Min, Weinberg Michael V, Wells Lance, Hoover Timothy R
Department of Microbiology, University of Georgia, Athens, GA 30602, USA.
Proteomics. 2007 Jun;7(13):2240-5. doi: 10.1002/pmic.200600875.
Helicobacter pylori extracellular proteins are of interest because of possible roles in pathogenesis, host recognition, and vaccine development. We utilized a unique approach by growing two strains (including one nonsequenced strain) in a defined serum-free medium and directly analyzing the proteins present in the culture supernatants by LC-MS/MS. Over 125 proteins were identified in the extracellular proteomes of two H. pylori strains. Forty-five of these proteins were enriched in the extracellular fraction when compared to soluble cell-associated protein samples. Our analysis confirmed and expanded on the previously reported H. pylori extracellular proteome. Extracellular proteins of interest identified here included cag pathogenicity island protein Cag24 (CagD); proteases HP0657 and HP1012; a polysaccharide deacetylase, HP0310, possibly involved in the hydrolysis of acetyl groups from host N-acetylglucosamine residues or from residues on the cell surface; and HP0953, an uncharacterized protein that appears to be restricted to Helicobacter species that colonize the gastric mucosa. In addition, our analysis found eight previously unidentified outer membrane proteins and two lipoproteins that could be important cell surface proteins.
幽门螺杆菌细胞外蛋白因其在发病机制、宿主识别和疫苗开发中可能发挥的作用而备受关注。我们采用了一种独特的方法，即在特定的无血清培养基中培养两种菌株（包括一种未测序菌株），并通过液相色谱-串联质谱法（LC-MS/MS）直接分析培养上清液中存在的蛋白质。在两种幽门螺杆菌菌株的细胞外蛋白质组中鉴定出了超过125种蛋白质。与可溶性细胞相关蛋白质样品相比，其中45种蛋白质在细胞外部分富集。我们的分析证实并扩展了先前报道的幽门螺杆菌细胞外蛋白质组。此处鉴定出的有意义的细胞外蛋白质包括cag致病岛蛋白Cag24（CagD）；蛋白酶HP0657和HP1012；一种多糖脱乙酰酶HP0310，可能参与从宿主N-乙酰葡糖胺残基或细胞表面残基上水解乙酰基；以及HP0953，一种未表征的蛋白质，似乎仅限于定殖于胃黏膜的幽门螺杆菌物种。此外，我们的分析发现了八种先前未鉴定的外膜蛋白和两种脂蛋白，它们可能是重要的细胞表面蛋白。
