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• W2337649133 abstract "We have recently shown that synthetic peptides of the effector domain of the low-molecular-mass GTP-binding protein Rab3 stimulate inositol 1,4,5-trisphosphate production in various permeabilized cells. To investigate the mechanism of the peptide-induced activation of phospholipase C (PLC) and to identify the PLC isoenzyme(s) targeted by these peptides, isolated pancreatic acinar membranes and cytosol were preincubated with anti-PLC antibodies before examination of PLC activity in response to the Rab3B/D effector-domain peptide (VSTVGIDFKVKTVYRH, peptide P1). Western blot analysis revealed the presence of PLC-β1, -β3, -γ1 and -δ1 in membrane and cytosolic fractions. P1 stimulated PLC activity in both membrane and cytosolic fractions. Anti-(PLC-β1) antibody inhibited P1-induced PLC activity in both subcellular fractions almost completely. Moreover, P1-induced amylase release in digitonin-permeabilized pancreatic acini was also inhibited. Other immunoneutralizing anti-PLC antibodies had no effect, suggesting that P1 activates PLC-β1 but not PLC-β3, -γ1 or -δ1. P1 also activated recombinant PLC-β1, indicating direct activation of PLC-β1 by Rab3 effector-domain peptides. To investigate further the structure–function relationship of the peptides, truncated peptides of P1 were tested for their ability to activate PLC in isolated pancreatic acinar membranes and to stimulate amylase release from digitonin-permeabilized pancreatic acini. Peptides containing a BXBXXXB(B) motif (where B represents a basic residue and X any residue) [KVKTVYRH (EC50 of 1 nM to stimulate amylase release) ≈ TVGIDFKVKTVYRH &gt; TVGIDFKVKTVYR] were potent stimulators of amylase release and PLC activity, whereas deletion of the C-terminus (VSTVGIDF), of the two basic C-terminal amino acid residues (VSTVGIDFKVKTVY and KVKTVY), or destruction of the BXB motif (VKTVYR) resulted in inactive peptides. In conclusion, the results of the present study show that short peptides containing a BXBXXXB motif represent promising pharmacological agents to activate the PLC-β1 isoenzyme." @default.
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• W2337649133 date "1997-09-15" @default.
• W2337649133 modified "2023-10-18" @default.
• W2337649133 title "Synthetic peptides containing a BXBXXXB(B) motif activate phospholipase C-<i>β</i>1" @default.
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• W2337649133 doi "https://doi.org/10.1042/bj3260669" @default.
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