STRUCTURAL STUDIES OF PHOSPHATASES, AND PARVALBUMINS

磷酸酶和小清蛋白的结构研究

• 批准号: 8361652
• 负责人: JOHN J TANNER
• 金额: $ 1.65万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361652
• 关键词: Acid Phosphatase; Active Sites; Affinity; Alkaline Earth Metals; Bacillus anthracis; Binding; Binding Sites; C-terminal; EF Hand Motifs; Enzymes; Exhibits; Family; Francisella tularensis; Funding; Grant; Haemophilus influenzae; Ions; Isoelectric Point; Membrane; Metal Binding Site; Metal Ion Binding; Metalloproteins; N-terminal; National Center for Research Resources; Nontypable Haemophilus influenza; Parvalbumins; Phosphoric Monoester Hydrolases; Principal Investigator; Protein Isoforms; Proteins; Recombinant Vaccines; Research; Research Infrastructure; Resources; Role; Source; Structure; United States National Institutes of Health; Vaccines; Variant; Virulence; base; cost; design; immunogenicity; magnesium ion; mutant; pathogenic bacteria; polypeptide; structural biology; vaccine development

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Two metalloproteins containing alkaline earth metal binding sites were investigated in this project: class C acid phosphatases and parvalbumins. Class C acid phosphatases (CCAPs) are outer membrane acid phosphatases that contain four essential Asp residues imbedded in the bipartite sequence motif of [IV]-[VAL]-D-[IL]-D-E-T-[VM]-L-X-[NT]-X(2)-Y in the N-terminal half of the polypeptide chain and [IV]-[LM]-X(2)-G-D-[NT]-L-X-D-F in the C-terminal half. The active site features a magnesium ion bound to three of the conserved Asp residues. Because of their localization to the bacterial outer membrane, CCAPs are potential candidates for vaccine development, and some progress has been made toward creating a vaccine against nontypeable Haemophilus influenzae using catalytically inactive mutants of the H. influenzae CCAP. We are studying the structure, function, and immunogenicity of CCAPs from pathogenic bacteria including H. influenzae, F. tularensis, and B. anthracis in order to understand the role of these enzymes in virulence and to provide a platform for the design of inactive mutants with enhanced immunogenicity for use in recombinant vaccines. The second aspect of this project focuses on parvalbumins, which are small (Mr=12,000), vertebrate-specific EF-hand proteins. The parvalbumin family includes alpha and beta sublineages, distinguished by isoelectric point and lineage-specific sequence differences. Despite the general similarity of their metal-ion binding sites and overall fold, parvalbumins exhibit broad variations in divalent ion affinity. We are exploring the physical and structural basis for these differences, using specific parvalbumin isoforms.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 在该项目中研究了两个含有碱性土金属结合位点的金属蛋白：C类酸性磷酸酶和白细胞蛋白酶。 C类酸性磷酸酶（CCAP）是外膜酸性磷酸酶，其中包含[IV] - [val] -d- [il] -d-e-t- t- [vm] -l-x- x- [nt]的两部分序列基序中嵌入的四个必不可少的ASP残基。 -x（2）-y多肽链的N末端一半和[iv] - [lm] -x（2）-g-d- [nt] -l-x-d-f在C末端的一半中。 活性位点具有与三个保守的ASP残基结合的镁离子。由于CCAP的本地化为细菌外膜，因此是疫苗发育的潜在候选者，并且已经采取了一些进展，用于使用催化型型流感的h. th. h. h. toply ccap的催化性突变体对不可抑制的嗜血性流感疫苗进行疫苗。 我们正在研究CCAP的结构，功能和免疫原性，包括流感流感，F。tularensis和B.炭疽病，以了解这些酶在毒力中的作用，并为无效突变体设计提供平台具有增强的免疫原性用于重组疫苗。 该项目的第二个方面的重点是小白蛋白，这些小蛋白小（MR = 12,000），脊椎动物特异性的EF手蛋白。 白蛋白蛋白家族包括α和βsublineages，由等电点和谱系特异性序列差异区别。 尽管其金属离子结合位点和整体折叠的一般相似性，但白蛋白蛋白在二价离子亲和力中表现出较大的变化。我们正在使用特定的白细胞蛋白同工型探索这些差异的物理和结构基础。