Cytosolic rat brain synapsin I is a diacylglycerol kinase.
The phosphorylation of diacylglycerol (DG), a reaction catalyzed by DG kinase, may be critical in the termination of effector-induced signals mediated by protein kinase C. Synapsin I is a principal target of intracellular protein kinases and is thought to be involved in the release of neurotransmitter from axon terminals. We present several lines of evidence which indicate that rat brain synapsin, in addition to this role, may function as a DG kinase. Purified rat brain DG kinase was digested with trypsin, which produced three major fragments whose sequence was identical to three regions in synapsin I. Using a rabbit anti-synapsin polyclonal antiserum, the elution profile of synapsin immunoreactivity coincided exactly with that of DG kinase activity in column fractions from the final step in the DG kinase purification procedure. As is the case with synapsin, the purified enzyme was a strongly basic protein with an isoelectric point greater than 10.0. Finally, incubating the DG kinase with highly purified bacterial collagenase, an enzyme that partially degrades the proline- and glycine-rich synapsin, resulted in the simultaneous loss of DG kinase activity and synapsin immunoreactivity. We conclude that cytosolic rat brain synapsin is capable of functioning as a DG kinase.
Kahn
D W
DW
Department of Cell Biology, Glaxo Inc., Research Triangle Park, NC 27709.
Besterman
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D002850:Chromatography, Gel; D003600:Cytosol / Q000201:enzymology; D019852:Diacylglycerol Kinase; D005260:Female; D008565:Membrane Proteins / Q000302:isolation & purification*; D003012:Microbial Collagenase / Q000378:metabolism; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010770:Phosphotransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D016704:Synapsins
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D016704:Synapsins; D010770:Phosphotransferases; D019852:Diacylglycerol Kinase; D003012:Microbial Collagenase