Gaf-1, a gamma -SNAP-binding protein associated with the mitochondria.
The role of alpha/beta-SNAP (Soluble NSF Attachment Protein) in vesicular trafficking is well established; however, the function of the ubiquitously expressed gamma-SNAP remains unclear. To further characterize the cellular role of this enigmatic protein, a two-hybrid screen was used to identify new, gamma-SNAP-binding proteins and to uncover potentially novel functions ... for gamma-SNAP. One such SNAP-binding protein, termed Gaf-1 (gamma-SNAP associate factor-1) specifically binds gamma- but not alpha-SNAP. The full-length Gaf-1 (75 kDa) is ubiquitously expressed and is found stoichiometrically associated with gamma-SNAP in cellular extracts. This binding is distinct from other SNAP interactions since no alpha-SNAP or NSF coprecipitated with Gaf-1. Subcellular fractionation and immunofluorescence analysis show that Gaf-1 is peripherally associated with the outer mitochondrial membrane. Only a fraction of gamma-SNAP was mitochondrial with the balance being either cytosolic or associated with other membrane fractions. GFP-gamma-SNAP and the C-terminal domain of Gaf-1 both show a reticular distribution in HEK-293 cells. This reticular structure colocalizes with Gaf-1 and mitochondria as well as with microtubules but not with other cytoskeletal elements. These data identify a class of gamma-SNAP interactions that is distinct from other members of the SNAP family and point to a potential role for gamma-SNAP in mitochondrial dynamics.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Carrier Proteins, Cell Line, Cytosol, Humans, Intracellular Membranes, Kinetics, Membrane Proteins, Mitochondria, Mitochondrial Proteins, Recombinant Proteins, Saccharomyces cerevisiae, Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins, Substrate Specificity, Transfection, Tubulin, Vesicular Transport Proteins
Adaptor Proteins, Signal Transducing, Carrier Proteins, Cell Line, Cytosol, Humans, Intracellular Membranes, Kinetics, Membrane Proteins, Mitochondria, Mitochondrial Proteins, Recombinant Proteins, Saccharomyces cerevisiae, Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins, Substrate Specificity, Transfection, Tubulin, Vesicular Transport Proteins
J. Biol. Chem.
Date: Apr. 20, 2001
PubMed ID: 11278501
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