Serum Proteins Influencing Helicobacter pylori Growth

描述(申请人提供)：幽门螺杆菌继续困扰着世界上很大一部分人口，导致从胃炎、溃疡到癌症等各种疾病。这种微生物在没有血清的情况下仍然很难培养，而且人们对有利于幽门螺杆菌生长的血清成分知之甚少。我们的初步数据表明，来自人或牛血清的铜蓝蛋白(Cp)和至少一种其他未知的血清蛋白负责促进生长。我们假设cp和另一种蛋白质协同作用，为幽门螺杆菌提供铁或保护细菌免受氧化应激。这项建议的具体目的是： 1.证实铜蓝蛋白是促进幽门螺杆菌生长的血清蛋白之一。 2.确定铜蓝蛋白抑制幽门螺杆菌生长的机制。 3.确定促进幽门螺杆菌生长所需的另一个血清蛋白(S)。 将使用各种生化测试和铜蓝蛋白同系物的克隆来源来实现目标1。目标2将包括检查铁的摄取和对氧化应激的抵抗力 铜蓝蛋白。我们将通过使用先进的质谱分析来鉴定血清部分中存在或不存在的蛋白质，然后测试纯化的蛋白质，从而实现目标3。 这项研究的结果可能会对幽门螺杆菌和其他挑剔细菌的研究产生广泛的影响。虽然许多研究都阐述了黏附因子对幽门螺杆菌定植的重要性，但对幽门螺杆菌生长和致病所需的其他宿主因子知之甚少。此外，许多其他致病物种仍然很难或不可能在体外培养。更全面地了解血清蛋白与细菌的相互作用也可能导致其他生物培养的进步。

DESCRIPTION (provided by applicant): Helicobacter pylori continues to afflict a large percentage of the human population worldwide, causing diseases ranging from gastritis and ulcers to cancer. This organism remains difficult to cultivate without serum, and little is known about serum components beneficial for H. pylori growth. Our preliminary data indicate that ceruloplasmin (cp) from human or bovine serum and at least one other unidentified serum protein are responsible for augmenting growth. We hypothesize that cp and another protein synergize to supply iron to H. pylori or protect the bacterium from oxidative stress. The specific aims of this proposal are to: 1. Confirm the identity of ceruloplasmin as one of the serum proteins augmenting H. pylori growth. 2. Determine the mechanism of ceruloplasmin effect on H. pylori growth. 3. Identify the other serum protein(s) required for augmentation of H. pylori growth. Various biochemical tests and cloned sources of ceruloplasmin homologs will be used to achieve aim 1. Aim 2 will involve examination of iron uptake and resistance to oxidative stress in the presence of ceruloplasmin. We will accomplish aim 3 by using advanced mass spectroscopic analysis to identify proteins present in or absent from serum fractions, followed by testing of purified proteins. Results from this study may have wide-ranging impacts on the study of H. pylori and other fastidious bacteria. Although many studies have addressed adhesion factors important for colonization, little is known about other host factors required for H. pylori growth and pathogenesis. Furthermore, numerous other pathogenic species remain difficult or impossible to cultivate in vitro. A more complete understanding of serum proteins interacting with bacteria could lead to advancements in cultivation of other organisms as well.