Direct detection of glycogenin reaction products during glycogen initiation

Thomas Hurley, Chad Walls, John R. Bennett, Peter Roach, Mu Wang

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Glycogenin initiates glycogen synthesis in an autocatalytic reaction in which individual glucose residues are covalently linked to Tyrosine 194 in order to form a short priming chain of glucose residues that is a substrate for glycogen synthase which, combined with the branching enzyme, catalyzes the bulk synthesis of glycogen. We sought to develop a new enzymatic assay to better characterize both the chemical and enzymatic characteristics of this unusual reaction. By directly detecting the reaction products using electrospray mass spectrometry this procedure permits both the visualization of the intact individual reaction species produced as a function of time and quantitation of the levels of each of species. The quantitation of the reaction agrees well with previous measurements of both catalytic rate and the change in rate as a function of average glucosylation. The results from this assay provide new insight into the mechanism by which glycogenin catalyzes the initiation reaction.

Original languageEnglish
Pages (from-to)374-378
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume348
Issue number2
DOIs
StatePublished - Sep 22 2006

Fingerprint

Glycogen
Reaction products
Assays
1,4-alpha-Glucan Branching Enzyme
Glucose
Glycogen Synthase
Enzyme Assays
Mass spectrometry
Tyrosine
Mass Spectrometry
Visualization
Substrates
glycogenin

Keywords

  • Glycogen synthesis
  • Glycosyltransferase
  • Mass spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Direct detection of glycogenin reaction products during glycogen initiation. / Hurley, Thomas; Walls, Chad; Bennett, John R.; Roach, Peter; Wang, Mu.

In: Biochemical and Biophysical Research Communications, Vol. 348, No. 2, 22.09.2006, p. 374-378.

Research output: Contribution to journalArticle

@article{258fc4655d6b4364a0b52f35efe39d7e,
title = "Direct detection of glycogenin reaction products during glycogen initiation",
abstract = "Glycogenin initiates glycogen synthesis in an autocatalytic reaction in which individual glucose residues are covalently linked to Tyrosine 194 in order to form a short priming chain of glucose residues that is a substrate for glycogen synthase which, combined with the branching enzyme, catalyzes the bulk synthesis of glycogen. We sought to develop a new enzymatic assay to better characterize both the chemical and enzymatic characteristics of this unusual reaction. By directly detecting the reaction products using electrospray mass spectrometry this procedure permits both the visualization of the intact individual reaction species produced as a function of time and quantitation of the levels of each of species. The quantitation of the reaction agrees well with previous measurements of both catalytic rate and the change in rate as a function of average glucosylation. The results from this assay provide new insight into the mechanism by which glycogenin catalyzes the initiation reaction.",
keywords = "Glycogen synthesis, Glycosyltransferase, Mass spectrometry",
author = "Thomas Hurley and Chad Walls and Bennett, {John R.} and Peter Roach and Mu Wang",
year = "2006",
month = "9",
day = "22",
doi = "10.1016/j.bbrc.2006.07.106",
language = "English",
volume = "348",
pages = "374--378",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Direct detection of glycogenin reaction products during glycogen initiation

AU - Hurley, Thomas

AU - Walls, Chad

AU - Bennett, John R.

AU - Roach, Peter

AU - Wang, Mu

PY - 2006/9/22

Y1 - 2006/9/22

N2 - Glycogenin initiates glycogen synthesis in an autocatalytic reaction in which individual glucose residues are covalently linked to Tyrosine 194 in order to form a short priming chain of glucose residues that is a substrate for glycogen synthase which, combined with the branching enzyme, catalyzes the bulk synthesis of glycogen. We sought to develop a new enzymatic assay to better characterize both the chemical and enzymatic characteristics of this unusual reaction. By directly detecting the reaction products using electrospray mass spectrometry this procedure permits both the visualization of the intact individual reaction species produced as a function of time and quantitation of the levels of each of species. The quantitation of the reaction agrees well with previous measurements of both catalytic rate and the change in rate as a function of average glucosylation. The results from this assay provide new insight into the mechanism by which glycogenin catalyzes the initiation reaction.

AB - Glycogenin initiates glycogen synthesis in an autocatalytic reaction in which individual glucose residues are covalently linked to Tyrosine 194 in order to form a short priming chain of glucose residues that is a substrate for glycogen synthase which, combined with the branching enzyme, catalyzes the bulk synthesis of glycogen. We sought to develop a new enzymatic assay to better characterize both the chemical and enzymatic characteristics of this unusual reaction. By directly detecting the reaction products using electrospray mass spectrometry this procedure permits both the visualization of the intact individual reaction species produced as a function of time and quantitation of the levels of each of species. The quantitation of the reaction agrees well with previous measurements of both catalytic rate and the change in rate as a function of average glucosylation. The results from this assay provide new insight into the mechanism by which glycogenin catalyzes the initiation reaction.

KW - Glycogen synthesis

KW - Glycosyltransferase

KW - Mass spectrometry

UR - http://www.scopus.com/inward/record.url?scp=33746907025&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33746907025&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2006.07.106

DO - 10.1016/j.bbrc.2006.07.106

M3 - Article

VL - 348

SP - 374

EP - 378

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -