C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases

Yuh Ping Chong, Andrew S. Chan, Khai Chew Chan, Nicholas A. Williamson, Edwina C. Lerner, Thomas E. Smithgall, Jeffrey D. Bjorge, Donald J. Fujita, Anthony W. Purcell, Glen Scholz, Terrence D. Mulhern, Heung Chin Cheng

Research output: Contribution to journalArticleResearchpeer-review

28 Citations (Scopus)

Abstract

The Src family of protein kinases (SFKs) mediates mitogenic signal transduction, and constitutive SFK activation is associated with tumorigenesis. To prevent constitutive SFK activation, the catalytic activity of SFKs in normal mammalian cells is suppressed mainly by two inhibitors called C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK), which inactivate SFKs by phosphorylating a consensus tyrosine near the C terminus of SFKs (Y T ). The phosphorylated Y T intramolecularly binds to the SH2 domain of SFKs. This interaction, known as pYT/SH2 interaction, together with binding between the SH2 kinase linker and the SH3 domain of SFKs (linker/SH3 interaction) stabilizes SFKs in a "closed" inactive conformation. We previously discovered an alternative mechanism CHK employs to inhibit SFKs. This mechanism, referred to as the non-catalytic inhibitory mechanism, involves tight binding of CHK to SFKs; the binding alone is sufficient to inhibit SFKs. Herein, we constructed multiple active conformations of an SFK member, Hck, by systematically disrupting the two inhibitory interactions. We found that CHK employs the non-catalytic mechanism to inactivate these active conformations of Hck. However, CHK does not bind Hck when it adopts the inactive conformation in which both inhibitory interactions are intact. These data indicate that binding of CHK to SFKs via the non-catalytic mechanism is governed by the conformations of SFKs. Although CSK is also an inhibitor of SFKs, it does not inhibit SFKs by a similar non-catalytic mechanism. Thus, the non-catalytic inhibitory mechanism is a unique property of CHK that allows it to down-regulate multiple active conformations of SFKs.

Original languageEnglish
Pages (from-to)32988-32999
Number of pages12
JournalJournal of Biological Chemistry
Volume281
Issue number44
DOIs
Publication statusPublished - 3 Nov 2006

Cite this

Chong, Y. P., Chan, A. S., Chan, K. C., Williamson, N. A., Lerner, E. C., Smithgall, T. E., ... Cheng, H. C. (2006). C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases. Journal of Biological Chemistry, 281(44), 32988-32999. https://doi.org/10.1074/jbc.M602951200
Chong, Yuh Ping ; Chan, Andrew S. ; Chan, Khai Chew ; Williamson, Nicholas A. ; Lerner, Edwina C. ; Smithgall, Thomas E. ; Bjorge, Jeffrey D. ; Fujita, Donald J. ; Purcell, Anthony W. ; Scholz, Glen ; Mulhern, Terrence D. ; Cheng, Heung Chin. / C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 44. pp. 32988-32999.
@article{08a20a8dc5c54572ba6548b696be0dde,
title = "C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases",
abstract = "The Src family of protein kinases (SFKs) mediates mitogenic signal transduction, and constitutive SFK activation is associated with tumorigenesis. To prevent constitutive SFK activation, the catalytic activity of SFKs in normal mammalian cells is suppressed mainly by two inhibitors called C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK), which inactivate SFKs by phosphorylating a consensus tyrosine near the C terminus of SFKs (Y T ). The phosphorylated Y T intramolecularly binds to the SH2 domain of SFKs. This interaction, known as pYT/SH2 interaction, together with binding between the SH2 kinase linker and the SH3 domain of SFKs (linker/SH3 interaction) stabilizes SFKs in a {"}closed{"} inactive conformation. We previously discovered an alternative mechanism CHK employs to inhibit SFKs. This mechanism, referred to as the non-catalytic inhibitory mechanism, involves tight binding of CHK to SFKs; the binding alone is sufficient to inhibit SFKs. Herein, we constructed multiple active conformations of an SFK member, Hck, by systematically disrupting the two inhibitory interactions. We found that CHK employs the non-catalytic mechanism to inactivate these active conformations of Hck. However, CHK does not bind Hck when it adopts the inactive conformation in which both inhibitory interactions are intact. These data indicate that binding of CHK to SFKs via the non-catalytic mechanism is governed by the conformations of SFKs. Although CSK is also an inhibitor of SFKs, it does not inhibit SFKs by a similar non-catalytic mechanism. Thus, the non-catalytic inhibitory mechanism is a unique property of CHK that allows it to down-regulate multiple active conformations of SFKs.",
author = "Chong, {Yuh Ping} and Chan, {Andrew S.} and Chan, {Khai Chew} and Williamson, {Nicholas A.} and Lerner, {Edwina C.} and Smithgall, {Thomas E.} and Bjorge, {Jeffrey D.} and Fujita, {Donald J.} and Purcell, {Anthony W.} and Glen Scholz and Mulhern, {Terrence D.} and Cheng, {Heung Chin}",
year = "2006",
month = "11",
day = "3",
doi = "10.1074/jbc.M602951200",
language = "English",
volume = "281",
pages = "32988--32999",
journal = "Journal of Biological Chemistry",
issn = "1083-351X",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "44",

}

Chong, YP, Chan, AS, Chan, KC, Williamson, NA, Lerner, EC, Smithgall, TE, Bjorge, JD, Fujita, DJ, Purcell, AW, Scholz, G, Mulhern, TD & Cheng, HC 2006, 'C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases', Journal of Biological Chemistry, vol. 281, no. 44, pp. 32988-32999. https://doi.org/10.1074/jbc.M602951200

C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases. / Chong, Yuh Ping; Chan, Andrew S.; Chan, Khai Chew; Williamson, Nicholas A.; Lerner, Edwina C.; Smithgall, Thomas E.; Bjorge, Jeffrey D.; Fujita, Donald J.; Purcell, Anthony W.; Scholz, Glen; Mulhern, Terrence D.; Cheng, Heung Chin.

In: Journal of Biological Chemistry, Vol. 281, No. 44, 03.11.2006, p. 32988-32999.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - C-terminal Src kinase-homologous kinase (CHK), a unique inhibitor inactivating multiple active conformations of Src family tyrosine kinases

AU - Chong, Yuh Ping

AU - Chan, Andrew S.

AU - Chan, Khai Chew

AU - Williamson, Nicholas A.

AU - Lerner, Edwina C.

AU - Smithgall, Thomas E.

AU - Bjorge, Jeffrey D.

AU - Fujita, Donald J.

AU - Purcell, Anthony W.

AU - Scholz, Glen

AU - Mulhern, Terrence D.

AU - Cheng, Heung Chin

PY - 2006/11/3

Y1 - 2006/11/3

N2 - The Src family of protein kinases (SFKs) mediates mitogenic signal transduction, and constitutive SFK activation is associated with tumorigenesis. To prevent constitutive SFK activation, the catalytic activity of SFKs in normal mammalian cells is suppressed mainly by two inhibitors called C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK), which inactivate SFKs by phosphorylating a consensus tyrosine near the C terminus of SFKs (Y T ). The phosphorylated Y T intramolecularly binds to the SH2 domain of SFKs. This interaction, known as pYT/SH2 interaction, together with binding between the SH2 kinase linker and the SH3 domain of SFKs (linker/SH3 interaction) stabilizes SFKs in a "closed" inactive conformation. We previously discovered an alternative mechanism CHK employs to inhibit SFKs. This mechanism, referred to as the non-catalytic inhibitory mechanism, involves tight binding of CHK to SFKs; the binding alone is sufficient to inhibit SFKs. Herein, we constructed multiple active conformations of an SFK member, Hck, by systematically disrupting the two inhibitory interactions. We found that CHK employs the non-catalytic mechanism to inactivate these active conformations of Hck. However, CHK does not bind Hck when it adopts the inactive conformation in which both inhibitory interactions are intact. These data indicate that binding of CHK to SFKs via the non-catalytic mechanism is governed by the conformations of SFKs. Although CSK is also an inhibitor of SFKs, it does not inhibit SFKs by a similar non-catalytic mechanism. Thus, the non-catalytic inhibitory mechanism is a unique property of CHK that allows it to down-regulate multiple active conformations of SFKs.

AB - The Src family of protein kinases (SFKs) mediates mitogenic signal transduction, and constitutive SFK activation is associated with tumorigenesis. To prevent constitutive SFK activation, the catalytic activity of SFKs in normal mammalian cells is suppressed mainly by two inhibitors called C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK), which inactivate SFKs by phosphorylating a consensus tyrosine near the C terminus of SFKs (Y T ). The phosphorylated Y T intramolecularly binds to the SH2 domain of SFKs. This interaction, known as pYT/SH2 interaction, together with binding between the SH2 kinase linker and the SH3 domain of SFKs (linker/SH3 interaction) stabilizes SFKs in a "closed" inactive conformation. We previously discovered an alternative mechanism CHK employs to inhibit SFKs. This mechanism, referred to as the non-catalytic inhibitory mechanism, involves tight binding of CHK to SFKs; the binding alone is sufficient to inhibit SFKs. Herein, we constructed multiple active conformations of an SFK member, Hck, by systematically disrupting the two inhibitory interactions. We found that CHK employs the non-catalytic mechanism to inactivate these active conformations of Hck. However, CHK does not bind Hck when it adopts the inactive conformation in which both inhibitory interactions are intact. These data indicate that binding of CHK to SFKs via the non-catalytic mechanism is governed by the conformations of SFKs. Although CSK is also an inhibitor of SFKs, it does not inhibit SFKs by a similar non-catalytic mechanism. Thus, the non-catalytic inhibitory mechanism is a unique property of CHK that allows it to down-regulate multiple active conformations of SFKs.

UR - http://www.scopus.com/inward/record.url?scp=33750214579&partnerID=8YFLogxK

U2 - 10.1074/jbc.M602951200

DO - 10.1074/jbc.M602951200

M3 - Article

VL - 281

SP - 32988

EP - 32999

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 1083-351X

IS - 44

ER -