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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 本项目研究了两种含碱土金属结合位点的金属蛋白：C类酸性磷酸酶和小清蛋白。C类酸性磷酸酶（CCAP）是外膜酸性磷酸酶，其含有四个必需的Asp残基，嵌入在多肽链的N-末端一半的[IV]-[瓦尔]-D-[IL]-D-E-T-[VM]-L-X-[NT]-X（2）-Y和C-末端一半的[IV]-[LM]-X（2）-G-D-[NT]-L-X-D-F的二分序列基序中。 活性位点的特点是镁离子结合到三个保守的天冬氨酸残基。由于它们定位于细菌外膜，CCAP是疫苗开发的潜在候选者，并且已经在使用H.流感CCAP。 我们正在研究病原菌包括H. influenzae、F.土拉热菌（tularensis）和B.炭疽杆菌中，以了解这些酶的毒力的作用，并提供一个平台，用于设计无活性的突变体与增强免疫原性用于重组疫苗。 该项目的第二个方面集中在小清蛋白，这是小的（Mr= 12，000），脊椎动物特异性EF-手蛋白。 小清蛋白家族包括α和β亚系，通过等电点和谱系特异性序列差异来区分。 尽管它们的金属离子结合位点和整体折叠的一般相似性，小清蛋白在二价离子亲和力方面表现出广泛的变化。我们正在探索这些差异的物理和结构基础，使用特定的小清蛋白亚型。