Unknown

Dataset Information

0

Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase.


ABSTRACT: PCTAIRE-1 (cyclin-dependent kinase [CDK] 16) is a highly conserved serine/threonine kinase that belongs to the CDK family of protein kinases. Little is known regarding PCTAIRE-1 regulation and function and no robust assay exists to assess PCTAIRE-1 activity mainly due to a lack of information regarding its preferred consensus motif and the lack of bona fide substrates. We used positional scanning peptide library technology and identified the substrate-specificity requirements of PCTAIRE-1 and subsequently elaborated a peptide substrate termed PCTAIRE-tide. Recombinant PCTAIRE-1 displayed vastly improved enzyme kinetics on PCTAIRE-tide compared to a widely used generic CDK substrate peptide. PCTAIRE-tide also greatly improved detection of endogenous PCTAIRE-1 activity. Similar to other CDKs, PCTAIRE-1 requires a proline residue immediately C-terminal to the phosphoacceptor site (+1) for optimal activity. PCTAIRE-1 has a unique preference for a basic residue at +4, but not at +3 position (a key characteristic for CDKs). We also demonstrate that PCTAIRE-1 binds to a novel cyclin family member, cyclin Y, which increased PCTAIRE-1 activity towards PCTAIRE-tide >100-fold. We hypothesised that cyclin Y binds and activates PCTAIRE-1 in a way similar to which cyclin A2 binds and activates CDK2. Point mutants of cyclin Y predicted to disrupt PCTAIRE-1-cyclin Y binding severely prevented complex formation and activation of PCTAIRE-1. We have identified PCTAIRE-tide as a powerful tool to study the regulation of PCTAIRE-1. Our understanding of the molecular interaction between PCTAIRE-1 and cyclin Y further facilitates future investigation of the functions of PCTAIRE-1 kinase.

SUBMITTER: Shehata SN 

PROVIDER: S-EPMC3590450 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature

altmetric image

Publications

Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase.

Shehata Saifeldin N SN   Hunter Roger W RW   Ohta Eriko E   Peggie Mark W MW   Lou Hua Jane HJ   Sicheri Frank F   Zeqiraj Elton E   Turk Benjamin E BE   Sakamoto Kei K  

Cellular signalling 20120711 11


PCTAIRE-1 (cyclin-dependent kinase [CDK] 16) is a highly conserved serine/threonine kinase that belongs to the CDK family of protein kinases. Little is known regarding PCTAIRE-1 regulation and function and no robust assay exists to assess PCTAIRE-1 activity mainly due to a lack of information regarding its preferred consensus motif and the lack of bona fide substrates. We used positional scanning peptide library technology and identified the substrate-specificity requirements of PCTAIRE-1 and su  ...[more]

Similar Datasets

| S-EPMC3272973 | biostudies-literature
| S-EPMC5317395 | biostudies-literature
| S-EPMC4140294 | biostudies-literature
| S-EPMC3059039 | biostudies-literature
| S-EPMC3778134 | biostudies-literature
| S-EPMC3973122 | biostudies-literature
| S-EPMC5209680 | biostudies-literature
| S-EPMC2263080 | biostudies-literature
| S-EPMC3425359 | biostudies-literature
| S-EPMC10327203 | biostudies-literature