SpyTag/SpyCatcher Cyclization Enhances the Thermostability of Firefly Luciferase

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2016 Sep 23

PMID 27658030

Citations 27

Authors

Meng Si

Qing Xu

Ling Jiang

He Huang

Affiliations

Soon will be listed here.

Abstract

SpyTag can spontaneously form a covalent isopeptide bond with its protein partner SpyCatcher. Firefly luciferase from Photinus pyralis was cyclized in vivo by fusing SpyCatcher at the N terminus and SpyTag at the C terminus. Circular LUC was more thermostable and alkali-tolerant than the wild type, without compromising the specific activity. Structural analysis indicated that the cyclized LUC increased the thermodynamic stability of the structure and remained more properly folded at high temperatures when compared with the wild type. We also prepared an N-terminally and C-terminally shortened form of the SpyCatcher protein and cyclization using this truncated form led to even more thermostability than the original form. Our findings suggest that cyclization with SpyTag and SpyCatcher is a promising and effective strategy to enhance thermostability of enzymes.

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