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

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Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase. / Shehata, Saifeldin N.; Hunter, Roger W.; Ohta, Eriko; Peggie, Mark W.; Lou, Hua Jane; Sicheri, Frank; Zeqiraj, Elton; Turk, Benjamin E.; Sakamoto, Kei.

In: Cellular Signalling, Vol. 24, No. 11, 01.11.2012, p. 2085-2094.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Shehata, SN, Hunter, RW, Ohta, E, Peggie, MW, Lou, HJ, Sicheri, F, Zeqiraj, E, Turk, BE & Sakamoto, K 2012, 'Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase', Cellular Signalling, vol. 24, no. 11, pp. 2085-2094. https://doi.org/10.1016/j.cellsig.2012.06.018

APA

Shehata, S. N., Hunter, R. W., Ohta, E., Peggie, M. W., Lou, H. J., Sicheri, F., Zeqiraj, E., Turk, B. E., & Sakamoto, K. (2012). Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase. Cellular Signalling, 24(11), 2085-2094. https://doi.org/10.1016/j.cellsig.2012.06.018

Vancouver

Shehata SN, Hunter RW, Ohta E, Peggie MW, Lou HJ, Sicheri F et al. Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase. Cellular Signalling. 2012 Nov 1;24(11):2085-2094. https://doi.org/10.1016/j.cellsig.2012.06.018

Author

Shehata, Saifeldin N. ; Hunter, Roger W. ; Ohta, Eriko ; Peggie, Mark W. ; Lou, Hua Jane ; Sicheri, Frank ; Zeqiraj, Elton ; Turk, Benjamin E. ; Sakamoto, Kei. / Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase. In: Cellular Signalling. 2012 ; Vol. 24, No. 11. pp. 2085-2094.

Bibtex

@article{b31acdb67d204ca183d57a083be2cc6c,
title = "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.",
keywords = "CDK16, Cell cycle, Cyclin-dependent kinase, PCTK1, Positional scanning peptide library, Proline-directed kinase",
author = "Shehata, {Saifeldin N.} and Hunter, {Roger W.} and Eriko Ohta and Peggie, {Mark W.} and Lou, {Hua Jane} and Frank Sicheri and Elton Zeqiraj and Turk, {Benjamin E.} and Kei Sakamoto",
year = "2012",
month = nov,
day = "1",
doi = "10.1016/j.cellsig.2012.06.018",
language = "English",
volume = "24",
pages = "2085--2094",
journal = "Cellular Signalling",
issn = "0898-6568",
publisher = "Elsevier",
number = "11",

}

RIS

TY - JOUR

T1 - Analysis of substrate specificity and cyclin Y binding of PCTAIRE-1 kinase

AU - Shehata, Saifeldin N.

AU - Hunter, Roger W.

AU - Ohta, Eriko

AU - Peggie, Mark W.

AU - Lou, Hua Jane

AU - Sicheri, Frank

AU - Zeqiraj, Elton

AU - Turk, Benjamin E.

AU - Sakamoto, Kei

PY - 2012/11/1

Y1 - 2012/11/1

N2 - 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.

AB - 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.

KW - CDK16

KW - Cell cycle

KW - Cyclin-dependent kinase

KW - PCTK1

KW - Positional scanning peptide library

KW - Proline-directed kinase

UR - http://www.scopus.com/inward/record.url?scp=84865339164&partnerID=8YFLogxK

U2 - 10.1016/j.cellsig.2012.06.018

DO - 10.1016/j.cellsig.2012.06.018

M3 - Journal article

C2 - 22796189

AN - SCOPUS:84865339164

VL - 24

SP - 2085

EP - 2094

JO - Cellular Signalling

JF - Cellular Signalling

SN - 0898-6568

IS - 11

ER -

ID: 239566412