Investigating Substrate Specificity of Sortases

研究分选酶的底物特异性

• 批准号: 7277275
• 负责人: Valerie A Villareal
• 金额: $ 2.96万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7277275
• 关键词: Amino Acid Motifs; Amino Acid Sequence; Amino Acids; Bacteria; Bacterial Adhesins; Bioinformatics; Cell Wall; Cell surface; Complement; Complex; Computer Analysis; Disease; Enzymes; Family; Fellowship; Goals; Gram-Positive Bacteria; Immune system; In Vitro; Individual; Infection; Mediating; Membrane; Monitor; NMR Spectroscopy; Names; Nosocomial Infections; Numbers; Osteomyelitis; Peptidoglycan; Peptidyltransferase; Pneumonia; Process; Proteins; Range; Reporter; Signal Transduction; Solutions; Sorting - Cell Movement; Staphylococcus aureus; Streptococcus pyogenes; Structure; Substrate Specificity; Tissues; Variant; Virulence Factors; Work; base; design; in vivo; inhibitor/antagonist; insight; leucylproline; member; microbial genome; mouse model; protein aminoacid sequence; research study; sortase; threonyl-glycine

DESCRIPTION (provided by applicant): Gram-positive bacteria are the causative agents of many nosocomial infections and diseases including osteomyelitis and pneumonia. The majority of Gram-positive bacteria harbor sortases, which are transpeptidases that recognize a short peptide sequence found near the C-terminus of proteins. Proteins harboring this recognition motif are subsequently anchored to the peptidoglycan and are required to establish infection by acting as adhesins or by mediating evasion from the host's immune system. One such sortase, SrtA, recognizes and processes the sequence, Leu-Pro-X-Thr-Gly motif in vitro. Bioinformatics analyses predict SrtA may recognize LPXTG variants as well as a 6th amino acid. Another family of sortases, subfamily-3, is also predicted to process similar recognition motifs. We propose to determine the substrate specificity of these sortases to understand in vivo recognition. We will also use NMR spectroscopy to solve the structures of the sortase-substrate complexes. Results from this project will afford details on how these transpeptidases function and will identify residues participating in recognition. This work will complement ongoing sortase inhibitor studies.

描述（由申请人提供）：革兰氏阳性细菌是许多医院感染和疾病（包括骨髓炎和肺炎）的病因。大多数革兰氏阳性细菌都携带排序酶，它们是肽酶，它们识别蛋白质C末端附近发现的短肽序列。随后将具有这种识别基序的蛋白质锚定在肽聚糖上，并被要求通过充当粘附素或介导宿主免疫系统的逃避来建立感染。一种这样的排序酶SRTA识别并处理序列的leu-pro-x-th-th-gly基序体外。生物信息学分析预测SRTA可能识别LPXTG变体以及第六氨基酸。也预计另一个类别酶Subfibily-3家族将处理类似的识别基序。我们建议确定这些排序酶的底物特异性，以了解体内识别。我们还将使用NMR光谱法来解决分子酶基底复合物的结构。该项目的结果将提供有关这些转肽酶如何发挥作用并确定参与认可的残留物的细节。这项工作将补充正在进行的排序酶抑制剂研究。