Generation of Ca2+-independent Sortase A Mutants with Enhanced Activity for Protein and Cell Surface Labeling

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Dec 5

PMID 29200433

Citations 24

Authors

Hee-Jin Jeong

Gita C Abhiraman

Craig M Story

Jessica R Ingram

Stephanie K Dougan

Affiliations

Soon will be listed here.

Abstract

Sortase A, a calcium-dependent transpeptidase derived from Staphylococcus aureus, is used in a broad range of applications, such as the conjugation of fluorescent dyes and other moieties to proteins or to the surface of eukaryotic cells. In vivo and cell-based applications of sortase have been somewhat limited by the large range of calcium concentrations, as well as by the often transient nature of protein-protein interactions in living systems. In order to use sortase A for cell labeling applications, we generated a new sortase A variant by combining multiple mutations to yield an enzyme that was both calcium-independent and highly active. This variant has enhanced activity for both N- and C-terminal labeling, as well as for cell surface modification under physiological conditions.

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