Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2

Zoe M. Goeckeler, Ruthann A. Masaracchia, Qi Zeng, Teng Leong Chew, Patricia Gallagher, Robert B. Wysolmerski

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

Phosphorylation of myosin II regulatory light chains (RLC) by Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) is a critical step in the initiation of smooth muscle and non-muscle cell contraction. Post-translational modifications to MLCK down-regulate enzyme activity, suppressing RLC phosphorylation, myosin II activation, and tension development. Here we report that PAK2, a member of the Rho family of GTPasedependent kinases, regulates isometric tension development and myosin II RLC phosphorylation in saponin permeabilized endothelial monolayers. PAK2 blunts tension development by 75% while inhibiting diphosphorylation of myosin II RLC. Cdc42-activated placenta and recombinant, constitutively active PAK2 phosphorylate MLCK in vitro with a stoichiometry of 1.71 ± 0.21 tool of PO4/mol of MLCK. This phosphorylation inhibits MLCK phosphorylation of myosin II RLC. PAK2 catalyzes MLCK phosphorylation on serine residues 439 and 991. Binding calmodulin to MLCK blocks phosphorylation of Ser-991 by PAK2. These results demonstrate that PAK2 can directly phosphorylate MLCK, inhibiting its activity and limiting the development of isometric tension.

Original languageEnglish
Pages (from-to)18366-18374
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number24
DOIs
StatePublished - Jun 16 2000

Fingerprint

p21-Activated Kinases
Myosin-Light-Chain Kinase
Phosphorylation
Myosin Type II
Myosin Light Chains
Calmodulin
Saponins
Enzyme activity
Post Translational Protein Processing
Stoichiometry
Serine
Placenta
Smooth Muscle
Muscle
Monolayers
Phosphotransferases
Down-Regulation
Chemical activation
Cells
Light

ASJC Scopus subject areas

  • Biochemistry

Cite this

Goeckeler, Z. M., Masaracchia, R. A., Zeng, Q., Chew, T. L., Gallagher, P., & Wysolmerski, R. B. (2000). Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2. Journal of Biological Chemistry, 275(24), 18366-18374. https://doi.org/10.1074/jbc.M001339200

Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2. / Goeckeler, Zoe M.; Masaracchia, Ruthann A.; Zeng, Qi; Chew, Teng Leong; Gallagher, Patricia; Wysolmerski, Robert B.

In: Journal of Biological Chemistry, Vol. 275, No. 24, 16.06.2000, p. 18366-18374.

Research output: Contribution to journalArticle

Goeckeler, ZM, Masaracchia, RA, Zeng, Q, Chew, TL, Gallagher, P & Wysolmerski, RB 2000, 'Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2', Journal of Biological Chemistry, vol. 275, no. 24, pp. 18366-18374. https://doi.org/10.1074/jbc.M001339200
Goeckeler ZM, Masaracchia RA, Zeng Q, Chew TL, Gallagher P, Wysolmerski RB. Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2. Journal of Biological Chemistry. 2000 Jun 16;275(24):18366-18374. https://doi.org/10.1074/jbc.M001339200
Goeckeler, Zoe M. ; Masaracchia, Ruthann A. ; Zeng, Qi ; Chew, Teng Leong ; Gallagher, Patricia ; Wysolmerski, Robert B. / Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 24. pp. 18366-18374.
@article{d53907a2546b4bc3a9744613e2591eb3,
title = "Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2",
abstract = "Phosphorylation of myosin II regulatory light chains (RLC) by Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) is a critical step in the initiation of smooth muscle and non-muscle cell contraction. Post-translational modifications to MLCK down-regulate enzyme activity, suppressing RLC phosphorylation, myosin II activation, and tension development. Here we report that PAK2, a member of the Rho family of GTPasedependent kinases, regulates isometric tension development and myosin II RLC phosphorylation in saponin permeabilized endothelial monolayers. PAK2 blunts tension development by 75{\%} while inhibiting diphosphorylation of myosin II RLC. Cdc42-activated placenta and recombinant, constitutively active PAK2 phosphorylate MLCK in vitro with a stoichiometry of 1.71 ± 0.21 tool of PO4/mol of MLCK. This phosphorylation inhibits MLCK phosphorylation of myosin II RLC. PAK2 catalyzes MLCK phosphorylation on serine residues 439 and 991. Binding calmodulin to MLCK blocks phosphorylation of Ser-991 by PAK2. These results demonstrate that PAK2 can directly phosphorylate MLCK, inhibiting its activity and limiting the development of isometric tension.",
author = "Goeckeler, {Zoe M.} and Masaracchia, {Ruthann A.} and Qi Zeng and Chew, {Teng Leong} and Patricia Gallagher and Wysolmerski, {Robert B.}",
year = "2000",
month = "6",
day = "16",
doi = "10.1074/jbc.M001339200",
language = "English",
volume = "275",
pages = "18366--18374",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "24",

}

TY - JOUR

T1 - Phosphorylation of myosin light chain kinase by p21-activated kinase PAK2

AU - Goeckeler, Zoe M.

AU - Masaracchia, Ruthann A.

AU - Zeng, Qi

AU - Chew, Teng Leong

AU - Gallagher, Patricia

AU - Wysolmerski, Robert B.

PY - 2000/6/16

Y1 - 2000/6/16

N2 - Phosphorylation of myosin II regulatory light chains (RLC) by Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) is a critical step in the initiation of smooth muscle and non-muscle cell contraction. Post-translational modifications to MLCK down-regulate enzyme activity, suppressing RLC phosphorylation, myosin II activation, and tension development. Here we report that PAK2, a member of the Rho family of GTPasedependent kinases, regulates isometric tension development and myosin II RLC phosphorylation in saponin permeabilized endothelial monolayers. PAK2 blunts tension development by 75% while inhibiting diphosphorylation of myosin II RLC. Cdc42-activated placenta and recombinant, constitutively active PAK2 phosphorylate MLCK in vitro with a stoichiometry of 1.71 ± 0.21 tool of PO4/mol of MLCK. This phosphorylation inhibits MLCK phosphorylation of myosin II RLC. PAK2 catalyzes MLCK phosphorylation on serine residues 439 and 991. Binding calmodulin to MLCK blocks phosphorylation of Ser-991 by PAK2. These results demonstrate that PAK2 can directly phosphorylate MLCK, inhibiting its activity and limiting the development of isometric tension.

AB - Phosphorylation of myosin II regulatory light chains (RLC) by Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) is a critical step in the initiation of smooth muscle and non-muscle cell contraction. Post-translational modifications to MLCK down-regulate enzyme activity, suppressing RLC phosphorylation, myosin II activation, and tension development. Here we report that PAK2, a member of the Rho family of GTPasedependent kinases, regulates isometric tension development and myosin II RLC phosphorylation in saponin permeabilized endothelial monolayers. PAK2 blunts tension development by 75% while inhibiting diphosphorylation of myosin II RLC. Cdc42-activated placenta and recombinant, constitutively active PAK2 phosphorylate MLCK in vitro with a stoichiometry of 1.71 ± 0.21 tool of PO4/mol of MLCK. This phosphorylation inhibits MLCK phosphorylation of myosin II RLC. PAK2 catalyzes MLCK phosphorylation on serine residues 439 and 991. Binding calmodulin to MLCK blocks phosphorylation of Ser-991 by PAK2. These results demonstrate that PAK2 can directly phosphorylate MLCK, inhibiting its activity and limiting the development of isometric tension.

UR - http://www.scopus.com/inward/record.url?scp=0034674408&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034674408&partnerID=8YFLogxK

U2 - 10.1074/jbc.M001339200

DO - 10.1074/jbc.M001339200

M3 - Article

VL - 275

SP - 18366

EP - 18374

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 24

ER -