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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Thaker, Tarjani M. Hamm, Heidi E. Sarwar, Maruf Preininger, Anita M. Iverson, T. M. |
| Description | Author Affiliation: Thaker TM ( From the Department of Biochemistry and.) |
| Abstract | Receptor-mediated activation of the G subunit of heterotrimeric G proteins requires allosteric communication between the receptor binding site and the guanine nucleotide binding site, which are separated by >30 ⠫. Structural changes in the allosteric network connecting these sites are predicted to be transient in the wild-type G subunit, making studies of these connections challenging. In the current work, site-directed mutants that alter the energy barriers between the activation states are used as tools to better understand the transient features of allosteric signaling in the G subunit. The observed differences in relative receptor affinity for intact G i1 subunits versus C-terminal G i1 peptides harboring the K345L mutation are consistent with this mutation modulating the allosteric network in the protein subunit. Measurement of nucleotide exchange rates, affinity for metarhodopsin II, and thermostability suggest that the K345L G i1 variant has reduced stability in both the GDP-bound and nucleotide-free states as compared with wild type but similar stability in the GTPγS-bound state. High resolution x-ray crystal structures reveal conformational changes accompanying the destabilization of the GDP-bound state. Of these, the conformation for Switch I was stabilized by an ionic interaction with the phosphate binding loop. Further site-directed mutagenesis suggests that this interaction between Switch I and the phosphate binding loop is important for receptor-mediated nucleotide exchange in the wild-type G i1 subunit. |
| ISSN | 00219258 |
| e-ISSN | 1083351X |
| Journal | Journal of Biological Chemistry |
| Issue Number | 16 |
| Volume Number | 289 |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date | 2014-04-18 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | GTP-Binding Protein Alpha Subunits, Gi-Go Chemistry Guanosine 5'-O-(3-Thiotriphosphate) Guanosine Diphosphate Allosteric Regulation Physiology Amino Acid Substitution Animals Crystallography, X-Ray Genetics Metabolism Mutation, Missense Protein Structure, Secondary Research Support, N.I.H., Extramural Biochemistry Molecular Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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