Engineering active lysostaphin variants that incorporate noncanonical amino acids and characterizing the effects of site‐specific PEGylation

Author:

Batugal Troy1,Pendyala Geetanjali2,Tomasovic Luke2,Varner Chad2,Caplin Jeremy D.34,Page Alexander M.5,Davis Michelle5,Satola Sarah W.5,García Andrés J.34,Kane Ravi S.23ORCID

Affiliation:

1. School of Materials Science and Engineering Georgia Institute of Technology Atlanta Georgia USA

2. School of Chemical & Biomolecular Engineering Georgia Institute of Technology Atlanta Georgia USA

3. Petit Institute for Bioengineering and Bioscience Georgia Institute of Technology Atlanta Georgia USA

4. Woodruff School of Mechanical Engineering Georgia Institute of Technology Atlanta Georgia USA

5. Department of Medicine, Division of Infectious Diseases and Emory Investigational Clinical Microbiology Core Emory University School of Medicine Atlanta Georgia USA

Abstract

AbstractWe describe a facile strategy to identify sites for the incorporation of noncanonical amino acids into lysostaphin—an enzyme that degrades the cell wall of Staphylococcus aureus—while retaining stapholytic activity. We used this strategy to generate active variants of lysostaphin incorporating para‐azidophenylalanine. The incorporation of this “reactive handle” enabled the orthogonal site‐specific modification of the enzyme variants with polyethylene glycol (PEG) using copper‐free click cycloaddition. PEGylated lysostaphin variants could retain their stapholytic activity, with the extent of retention depending on the site of modification and the PEG molecular weight. The site‐specific modification of lysostaphin could be useful not only for PEGylation to improve biocompatibility but also for the incorporation of the enzyme into hydrogels and other biomaterials and for studies of protein structure and dynamics. Moreover, the approach described herein could be readily applied to identify suitable sites for the incorporation of reactive handles into other proteins of interest.

Funder

National Institutes of Health

Publisher

Wiley

Subject

Applied Microbiology and Biotechnology,Bioengineering,Biotechnology

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3