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Inhibition of ras-induced oocyte maturation by peptides from ras-p21 and GTPase activating protein (GAP) identified as being effector domains from molecular dynamics calculations
Journal article   Peer reviewed

Inhibition of ras-induced oocyte maturation by peptides from ras-p21 and GTPase activating protein (GAP) identified as being effector domains from molecular dynamics calculations

Fred K Friedman, Lyndon Chie, Denise Chung, Richard Robinson, Paul Brandt-Rauf, Ziro Yamaizumi and Matthew R Pincus
Journal of protein chemistry, v 21(5), pp 361-366
Jul 2002
PMID: 12206510

Abstract

Amino Acid Sequence Amino Acid Substitution Animals Binding Sites Dose-Response Relationship, Drug Enzyme Inhibitors - chemistry Enzyme Inhibitors - pharmacology Escherichia coli GTPase-Activating Proteins - chemistry GTPase-Activating Proteins - pharmacology Insulin - pharmacology Molecular Sequence Data Oncogene Protein p21(ras) - antagonists & inhibitors Oncogene Protein p21(ras) - chemistry Oncogene Protein p21(ras) - metabolism Oncogene Protein p21(ras) - pharmacology Oocytes - drug effects Oocytes - growth & development Peptide Fragments - chemistry Peptide Fragments - pharmacology Progesterone - pharmacology Protein Structure, Tertiary Proto-Oncogene Proteins p21(ras) - chemistry Proto-Oncogene Proteins p21(ras) - pharmacology Signal Transduction - drug effects Time Factors Xenopus laevis
In the accompanying article, using molecular dynamics calculations, we found that the 66-77 and 122-138 domains in ras-p21 and the 821-827, 832-845, 917-924, 943-953, and 1003-1020 domains in GAP have different conformations in complexes of GAP with wild-type and oncogenic ras-p21. We have now synthesized peptides corresponding to each of these domains and coinjected them into oocytes with oncogenic p21, which induces oocyte maturation, or injected them into oocytes incubated with insulin that induces maturation by activating wild-type cellular ras-p21. We find that all of these peptides inhibit both agents but do not inhibit progesterone-induced maturation that occurs by a ras-independent pathway. The p21 66-77 and 122-138 peptides cause greater inhibition of oncogenic p21. On the other hand, the GAP 832-845 and 1003-1021 peptides inhibit insulin-induced maturation to a significantly greater extent. Since we have found that activated wild-type and oncogenic p21 activate downstream targets like raf differently, these GAP peptides may be useful probes for identifying elements unique to the wild-type ras-p21 pathway.

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Web of Science research areas
Biochemistry & Molecular Biology
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