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Identification and Characterization of Transducin Functional Cysteines, Lysines, and Acidic Residues by Group-Specific Labeling and Chemical Cross-Linking

  • José Bubis
  • Julio O. Ortiz
  • Carolina Möller
  • Enrique J. Milián

Abstract

Guanine nucleotide binding proteins or G-proteins function as molecular switches in a diverse set of signaling pathways by coupling seven-helix transmembrane receptors to specific intracellular effectors (Kaziro et al., 1991; Dohlman et al., 1991). G-proteins are heterotrimers composed of a-, ß-, and y-subunits. Activation of the appropiate receptor causes a GDP molecule bound to the resting form of a G-protein to be exchanged for GTP. As a consequence, the G-protein dissociates to form the a-subunit complexed to GTP, and the ßy-dimer. The GTP-bound conformation of the a-subunit is capable of activating or inhibiting a variety of downstream effectors including enzymes as well as ion channels (Birnbaumer, 1992; Hepler & Gilman, 1992; Simon et al., 1991). The released ßy-complex can itself activate or modulate some effectors (Logothetis et al., 1987; Tang et al., 1991; Katz et al., 1992). A GTPase-controlled timing mechanism inherent in all a-subunits and, in some cases, modulated by other proteins (Berstein et al., 1992; Arshaysky & Bownds, 1992), returns the GTP-activated a-subunit to the inactive GDP-bound conformation. The a-subunit complexed to GDP reassociates with the ßy-complex and forms again the hetero-trimer in its resting state. Conklin & Bourne (1993) proposed a structural model for a general G-protein a-subunit, on the basis of biochemical, immunologic, and molecular genetic observations. This model provided a blurred but revealing view of the orientation of membrane-bound Ga with regard to Gpy, receptors, and effectors.

Keywords

High Performance Liquid Chromatography Tryptic Peptide GTPase Activity Magnesium Acetate Guanine Nucleotide Binding Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • José Bubis
    • 1
  • Julio O. Ortiz
    • 1
  • Carolina Möller
    • 2
  • Enrique J. Milián
    • 1
  1. 1.Departamento de Biología CelularUniversidad Simon BolívarCaracasVenezuela
  2. 2.Departamento de QuímicaUniversidad Simón BolívarCaracasVenezuela

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