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一种非典型流感嗜血杆菌热核酸酶的特性。

Characterization of a nontypeable Haemophilus influenzae thermonuclease.

机构信息

Iowa Inflammation Program, The University of Iowa, Iowa City, Iowa City, IA, United States of America.

Department of Infectious Disease, University of Iowa Hospitals, Iowa City, IA, United States of America.

出版信息

PLoS One. 2018 May 10;13(5):e0197010. doi: 10.1371/journal.pone.0197010. eCollection 2018.

DOI:10.1371/journal.pone.0197010
PMID:29746527
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5944969/
Abstract

Nontypeable Haemophilus influenzae (NTHi) has been shown to form biofilms, comprised of extracellular DNA (eDNA), in the middle ear and bronchus during clinical infections. Studies in our laboratory have shown that NTHi possesses a homolog of Staphylococcus aureus thermonuclease (staphylococcal thermonuclease), NTHi nuclease (NTHi Nuc, HI_1296). This enzyme had similar size, heat stability, and divalent cation requirements to those of the staphylococcal homolog as determined by light scattering and circular dichroism spectroscopy. Small angle X-ray scattering (SAXS) analysis suggested an overall shape and substrate-binding site comparable to those of staphylococcal nuclease. However, NTHi Nuc was approximately 25-fold more active in fluorescence resonance energy transfer (FRET) activity assay than staphylococcal thermonuclease. Homology modeling implicates shorter NTHi Nuc loops near the active site for this enhanced activity.

摘要

无乳链球菌（NTHi）已被证明在临床感染期间在中耳和支气管中形成由细胞外 DNA（eDNA）组成的生物膜。我们实验室的研究表明，NTHi 拥有金黄色葡萄球菌热核酸酶（金黄色葡萄球菌热核酸酶）的同源物，NTHi 核酸酶（NTHi Nuc，HI_1296）。通过光散射和圆二色性光谱分析，该酶的大小、热稳定性和二价阳离子要求与金黄色葡萄球菌的同源物相似。小角度 X 射线散射（SAXS）分析表明，其整体形状和底物结合位点与金黄色葡萄球菌核酸酶相似。然而，NTHi Nuc 在荧光共振能量转移（FRET）活性测定中的活性比金黄色葡萄球菌热核酸酶高约 25 倍。同源建模表明，活性位点附近较短的 NTHi Nuc 环是这种增强活性的原因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/59ba676acefc/pone.0197010.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/d870ee135ad0/pone.0197010.g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/68f434d2e15e/pone.0197010.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/3a0a732e4f36/pone.0197010.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/d9b5bf4346d2/pone.0197010.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/c10c40612f92/pone.0197010.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/59ba676acefc/pone.0197010.g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/d870ee135ad0/pone.0197010.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/5a1a045bd4e6/pone.0197010.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/6d6365279cb9/pone.0197010.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/68f434d2e15e/pone.0197010.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/3a0a732e4f36/pone.0197010.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/d9b5bf4346d2/pone.0197010.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/c10c40612f92/pone.0197010.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d9ef/5944969/59ba676acefc/pone.0197010.g008.jpg

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