Role of dynamic interactions in effective signal transfer for Gβ stimulation of phospholipase C-β2

Elizabeth Buck, Peter Schatz, Suzanne Scarlata, Ravi Iyengar

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Heterotrimeric G protein subunits regulate their effectors by protein-protein interactions. The regions involved in these direct interactions have either signal transfer or general binding functions (Buck, E., Li, J., Chen, Y., Weng, G., Scarlata, S., and Iyengar, R. (1999) Science 283, 1332-1335). Although key determinants of signal transfer regions for G protein subunits have been identified, the mechanisms of signal transfer are not fully understood. We have used a combinatorial peptide approach to analyze one Gβ region, Gβ86-105, involved in signal transfer to the effector phospholipase C (PLC)-β2 to gain a more mechanistic understanding of Gβ/PLC-β2 signaling. Binding and functional studies with the combinatorial peptides on interaction with and stimulation/inhibition of phospholipase Cβ2 indicate that binding affinity can be resolved from EC50 for functional effects, such that peptides that have wild type binding affinities have 15- to 20-fold lower EC50 values. Although more potent, these peptides display a much lower extent of maximal stimulation. These peptides synergize with Gβγ or peptides encoding the second Gβ42-54 signal transfer region in maximally stimulating phospholipase C-β2. Other combinatorial peptides from the Gβ86-105 region that bind to PLC-β2 by themselves submaximally stimulate and extensively inhibit Gβγ stimulation of PLC-β2. The intrinsic stimulation function can be attributed to Arg-96 and Ser-97, the synergy function to Trp-99, and the binding affinity to Thr-87, Val-90, Pro-94, Arg-96, Ser-97, and Val-100. These results indicate that, even within signal transfer regions, residues involved in binding can be resolved from those involved in signal transfer and that signal transfer is likely to be achieved through dynamic rather than steady-state interactions.

Original languageEnglish
Pages (from-to)49707-49715
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number51
DOIs
StatePublished - 20 Dec 2002

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