Identification of the uridine 5'-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photoaffinity probe 5-azido-UDP-Glc

J Biol Chem. 1990 Mar 25;265(9):4782-4.

Abstract

Photoaffinity labeling of purified cellulose synthase with [beta-32P]5-azidouridine 5'-diphosphoglucose (UDP-Glc) has been used to identify the UDP-Glc binding subunit of the cellulose synthase from Acetobacter xylinum strain ATCC 53582. The results showed exclusive labeling of an 83-kDa polypeptide. Photoinsertion of [beta-32P]5-azido-UDP-Glc is stimulated by the cellulose synthase activator, bis-(3'----5') cyclic diguanylic acid. Addition of increasing amounts of UDP-Glc prevents photolabeling of the 83-kDa polypeptide. The reversible and photocatalyzed binding of this photoprobe also showed saturation kinetics. These studies demonstrate that the 83-kDa polypeptide is the catalytic subunit of the cellulose synthase in A. xylinum strain ATCC 53582.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Affinity Labels / metabolism*
  • Arabidopsis Proteins*
  • Autoradiography
  • Azides / metabolism*
  • Binding Sites
  • Cell Membrane / enzymology
  • Gluconacetobacter xylinus / enzymology*
  • Glucosyltransferases / isolation & purification
  • Glucosyltransferases / metabolism*
  • Macromolecular Substances
  • Molecular Weight
  • Phosphorus Radioisotopes
  • Uridine Diphosphate Glucose / analogs & derivatives
  • Uridine Diphosphate Glucose / metabolism*
  • Uridine Diphosphate Sugars / metabolism*

Substances

  • Affinity Labels
  • Arabidopsis Proteins
  • Azides
  • Macromolecular Substances
  • Phosphorus Radioisotopes
  • Uridine Diphosphate Sugars
  • 5-azidouridine 5'-diphosphoglucose
  • Glucosyltransferases
  • PRC1 protein, Arabidopsis
  • cellulose synthase (UDP-forming)
  • Uridine Diphosphate Glucose