Cloning, characterization, and expression of a novel GDP dissociation inhibitor isoform from skeletal muscle
UMass Chan Affiliations
Program in Molecular MedicineDocument Type
Journal ArticlePublication Date
1994-05-01Keywords
Amino Acid SequenceAnimals
Base Sequence
Blotting, Northern
Brain
Cell Line
Cloning, Molecular
DNA Primers
GTP-Binding Proteins
Glutathione Transferase
*Guanine Nucleotide Dissociation Inhibitors
Humans
Liver
Molecular Sequence Data
Muscles
Myocardium
Organ Specificity
Pancreas
Polymerase Chain Reaction
RNA
RNA, Messenger
Rats
Recombinant Fusion Proteins
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Species Specificity
Life Sciences
Medicine and Health Sciences
Metadata
Show full item recordAbstract
Cellular mechanisms for controlling membrane trafficking appear to involve small GTP-binding proteins such as the Rab proteins. Rab function is regulated by GDP dissociation inhibitor (GDI), which releases Rab proteins from membranes and inhibits GDP dissociation. Here we report the isolation of a full-length cDNA encoding a novel GDI isoform of 445 amino acids (GDI-2) with a deduced molecular weight of 50,649 from mouse skeletal muscle. Full-length and partial cDNA clones encoding a previously reported GDI protein (GDI-1) were also isolated from cDNA libraries prepared from rat brain and mouse skeletal muscle, respectively. The degree of deduced amino acid sequence identity between mouse GDI-2 and our mouse GDI-1 cDNA clone is 86%. Northern (RNA blot) analysis revealed that in human tissues, both GDI-1 and GDI-2 transcripts were abundant in brain, skeletal muscle, and pancreas but were weakly expressed in heart and liver. GDI-1 mRNA was expressed in kidney, whereas GDI-2 was almost absent, while in lung the relative amounts of these mRNA species were reversed. Specific antibodies against mouse GDI-1 and GDI-2 based on unique peptide sequences in the proteins were raised. Differentiation of 3T3-L1 fibroblasts into highly insulin-responsive adipocytes was accompanied by large increases in both mRNA and protein levels of GDI-1 and GDI-2. GDI-1 and GDI-2 expressed as glutathione S-transferase fusion proteins were both able to solubilize the membrane-bound forms of Rab4 and Rab5 in a GDP/GTP-dependent manner. Taken together, these data demonstrate that the protein products of at least two genes regulate the membrane dynamics of Rab proteins in mice.Source
Mol Cell Biol. 1994 May;14(5):3459-68.Permanent Link to this Item
http://hdl.handle.net/20.500.14038/38596PubMed ID
7513052Related Resources
Link to Article in PubMedCollections
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