Activity Detection of GalNAc Transferases by Protein-Based Fluorescence Sensors In Vivo

Lina Song; Collin Bachert; Adam D Linstedt

doi:10.1007/978-1-4939-6463-5_10

Activity Detection of GalNAc Transferases by Protein-Based Fluorescence Sensors In Vivo

Methods Mol Biol. 2016:1496:123-31. doi: 10.1007/978-1-4939-6463-5_10.

Authors

Lina Song¹, Collin Bachert¹, Adam D Linstedt²

Affiliations

¹ Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, 15213, USA.
² Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, 15213, USA. linstedt@cmu.edu.

Abstract

Mucin-type O-glycosylation occurring in the Golgi apparatus is an important protein posttranslational modification initiated by up to 20 GalNAc-transferase isozymes with largely distinct substrate specificities. Regulation of this enzyme family affects a vast array of proteins transiting the secretory pathway and misregulation causes human diseases. Here we describe the use of protein-based fluorescence sensors that traffic in the secretory pathway to monitor GalNAc-transferase activity in living cells. The sensors can either be "pan" or isozyme specific.

Keywords: Fluorescence-activating protein; Fluorescent biosensor; GalNAc transferase; O-glycosylation.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Biosensing Techniques / methods*
HEK293 Cells
Humans
Microscopy, Fluorescence / methods
N-Acetylgalactosaminyltransferases / genetics
N-Acetylgalactosaminyltransferases / metabolism*
Polypeptide N-acetylgalactosaminyltransferase

Substances

N-Acetylgalactosaminyltransferases

Abstract

Publication types

MeSH terms

Substances

Grants and funding