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| Content Provider | Journal of Biological Chemistry (JBC) |
|---|---|
| Author | Low, Boon Chuan Seow, Kah Tong Guy, Graeme R. |
| Abstract | We recently identified BNIP-2, a previously cloned Bcl-2- and E1B-associated protein, as a putative substrate of the FGF receptor tyrosine kinase and showed that it possesses GTPase-activating activity toward Cdc42 despite the lack of homology to previously described catalytic domains of GTPase-activating proteins (GAPs). BNIP-2 contains many arginine residues at the carboxyl terminus, which includes the region of homology to the noncatalytic domain of Cdc42GAP, termed BNIP-2 and Cdc42GAPhomology (BCH) domain. Using BNIP-2 glutathioneS-transferase recombinants, it was found that its BCH bound Cdc42, and contributed the GAP activity. This domain was predicted to fold into α-helical bundles similar to the topology of the catalytic GAP domain of Cdc42GAP. Alignment of exposed arginine residues in this domain helped to identify Arg-235 and Arg-238 as good candidates for catalysis. Arg-238 matched well to the arginine “finger” required for enhanced GTP hydrolysis in homodimerized Cdc42. Site-directed mutagenesis confirmed that an R235K or R238K mutation severely impaired the BNIP-2 GAP activity without affecting its binding to Cdc42. From deletion studies, a region adjacent to the arginine patch (288EYV290 on BNIP-2) and the Switch I and Rho family-specific “Insert” region on Cdc42 are involved in the binding. The results indicate that the BCH domain of BNIP-2 represents a novel GAP domain that employs an arginine patch motif similar to that of the Cdc42-homodimer. |
| Related Links | http://www.jbc.org/content/275/19/14415.abstract |
| Ending Page | 14422 |
| Starting Page | 14415 |
| Page Count | 8 |
| File Format | HTM / HTML PDF |
| ISSN | 00219258 |
| Journal | Journal of Biological Chemistry (JBC) |
| Issue Number | 19 |
| Volume Number | 275 |
| DOI | 10.1074/jbc.275.19.14415 |
| e-ISSN | 1083351X |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2000-05-12 |
| Access Restriction | Open |
| Subject Keyword | GTPase-activating protein (GAP) BNIP-2 and Cdc42GAP homology (BCH) Breakpoint cluster region homology (BH) GlutathioneS-transferase (GST) Guanosine 5′-3-O-(thio)triphosphate (GTPγS) Hemagglutinin (HA) Polyacrylamide gel electrophoresis (PAGE) MECHANISMS OF SIGNAL TRANSDUCTION |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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