Attenuation of GTPase activity of recombinant G(o) alpha by peptides representing sequence permutations of mastoparan.
There is convincing evidence that the cytoplasmic domains of multispanning receptors interact with guanine nucleotide-binding proteins (G proteins). What are the rules governing these interactions? In an attempt to answer this question, we focused our attention on mastoparan, an amphiphilic tetradecapeptide from wasp venom, and on nine of its variants, produced by sequence permutation, which have altered amphiphilicity or no amphiphilicity at all. Mastoparan enhances the GTPase activity of recombinant G(o) alpha 5-fold in phospholipid vesicles. Like mastoparan, four of the synthetic variants can form amphiphilic alpha-helices and two of them indeed stimulate the GTPase activity of the G protein, whereas the other two have no effect. This confirms that the activation of certain G proteins by a number of peptides is mainly due to their cationic amphiphilicity. However, this structural feature is clearly not sufficient. The relative orientation of the positively charged residues as well as that of the hydrophobic side chains appear to be of fundamental importance. The other five peptides are not amphiphilic and do not enhance the rate of GTP hydrolysis. Surprisingly, three of them almost completely inhibit the G protein's intrinsic GTPase activity. This finding is of interest because of the possible role differential regulation of G protein activity can play in cellular functions.
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Istituto Guido Donegani, Laboratorio di Biotecnologia, Monterotondo/Rome, Italy.
Wagner
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Crisari
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Camerini
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Tocchini Valentini
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GP
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D020558:GTP Phosphohydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D010455:Peptides; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014862:Wasp Venoms / Q000737:chemistry* / Q000494:pharmacology
D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D011994:Recombinant Proteins; D014862:Wasp Venoms; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C037500:mastoparan; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins