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An evaluation of strategies available for the identification of GTP-binding proteins required in intracellular signalling pathways.

Abstract：

:Strategies which can be used to elucidate the nature of a GTP-binding regulatory protein (G-protein) involved in an intracellular pathway of interest in the complex environment of the cell are described and evaluated. A desirable strategy is considered to be one in which the first stage indicates a requirement for one or more G-proteins, provides information on whether a monomeric, trimeric or other type of G-protein is involved, and gives some idea of the G-protein sub-class. In the second stage the specific G-protein involved is identified. Approaches available for investigations in the first stage include the use of analogues of GTP and GDP, AlF4-, inhibitors of G-protein isoprenylation, bacterial toxins which covalently modify G-proteins, and the introduction of a purified GDP dissociation inhibitor, GDP exchange and/or GTP-ase activating protein. Identification of the specific G-protein in the second stage can be achieved using anti G-protein antibodies, G-protein-or receptor-derived peptides, antisense G-protein RNA and over-expressed, constitutively-active or dominant-negative G-protein mutants. The correct interpretation of results obtained with GTP and GDP analogues and AlF4- in the first stage is complex and often difficult, and requires a thorough understanding of the functions and mechanisms of activation of G-proteins. Nevertheless, it is important to reach the correct conclusion at this stage since considerable time and expense are usually required for investigations in the second stage.

journal_name

Cell Signal

journal_title

Cellular signalling

authors

Barritt GJ,Gregory RB

doi

10.1016/s0898-6568(96)00131-3

subject

Has Abstract

pub_date

1997-05-01 00:00:00

pages

207-18

issue

3-4

eissn

0898-6568

issn

1873-3913

pii

S0898-6568(96)00131-3

journal_volume

9

pub_type

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