The Golgi matrix protein GM130: a specific interacting partner of the small GTPase rab1b

EMBO Rep. 2001 Apr;2(4):336-41. doi: 10.1093/embo-reports/kve065.

Abstract

To detect specific partners of the small Golgi-localized GTPase rab1b we generated rab1b mutants and used them as bait proteins in yeast two-hybrid screens. We isolated several specifically interacting clones. Two of them encode large protein fragments highly homologous to rat GM130 and to human Golgin95. The full-length human GM130 cDNA was cloned and its interaction with rab1b was characterized in detail by yeast two-hybrid and in vitro binding assays. Here we report for the first time that the rab1b protein interacts specifically with GM130 in a GTP-dependent manner and therefore needs the hypervariable regions of the N- and C-termini. We mapped the rab1b binding site of GM130 and provide evidence that it is different to the previously described p115 and Grasp65 binding sites of the GM130 protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoantigens
  • Binding Sites
  • Carrier Proteins / metabolism
  • Cell Line
  • Cloning, Molecular
  • DNA, Complementary / metabolism
  • Gene Library
  • Golgi Apparatus / metabolism*
  • Golgi Matrix Proteins
  • HeLa Cells
  • Humans
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Microscopy, Fluorescence
  • Models, Genetic
  • Molecular Sequence Data
  • Mutation
  • Plasmids / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Rats
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Transfection
  • Transformation, Genetic
  • Two-Hybrid System Techniques
  • Vesicular Transport Proteins*
  • beta-Galactosidase / metabolism
  • rab GTP-Binding Proteins / chemistry
  • rab GTP-Binding Proteins / metabolism*
  • rab1 GTP-Binding Proteins*

Substances

  • Autoantigens
  • Carrier Proteins
  • DNA, Complementary
  • GORASP1 protein, human
  • Golgi Matrix Proteins
  • Golgin subfamily A member 2
  • Gorasp1 protein, rat
  • Membrane Proteins
  • Recombinant Proteins
  • Vesicular Transport Proteins
  • vesicular transport factor p115
  • beta-Galactosidase
  • RAB1B protein, rat
  • rab GTP-Binding Proteins
  • rab1 GTP-Binding Proteins