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and
*Institute of Molecular and Cell Biology, Singapore 117609, Singapore; and
cDNA clones encoding a novel protein (VAMP5) homologous to
synaptobrevins/VAMPs are detected during database searches. The predicted 102-amino acid VAMP5 harbors a 23-residue hydrophobic region
near the carboxyl terminus and exhibits an overall amino acid identity
of 33% with synaptobrevin/VAMP1 and 2 and cellubrevin. Northern blot
analysis reveals that the mRNA for VAMP5 is preferentially expressed in
the skeletal muscle and heart, whereas significantly lower levels are
detected in several other tissues but not in the brain. During in vitro
differentiation (myogenesis) of C2C12 myoblasts into myotubes, the mRNA
level for VAMP5 is increased ~8- to 10-fold. Immunoblot
analysis using antibodies specific for VAMP5 shows that the protein
levels are also elevated ~6-fold during in vitro myogenesis of C2C12
cells. Indirect immunofluorescence microscopy and immunoelectron
microscopy reveal that VAMP5 is associated with the plasma membrane as
well as intracellular perinuclear and peripheral vesicular structures
of myotubes. Epitope-tagged versions of VAMP5 are similarly targeted to
the plasma membrane.
Centre for Microscopy and Microanalysis
and
Centre for Molecular and Cellular Biology,
Department of Physiology and Pharmacology, University of Queensland, St
Lucia, Queensland 4072, Australia
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