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Enzyme kinetics and interaction studies for human JNK1?1 and substrates activating transcription factor 2 (ATF2) and c-Jun N-terminal kinase (c-Jun).

ABSTRACT: c-Jun N-terminal kinase (JNK) is a stress signal transducer linked to cell death, and survival. JNK1 has been implicated in obesity, glucose intolerance, and insulin resistance. In this study we report the kinetic mechanism for JNK1?1 with transcription factors ATF2 and c-Jun along with interaction kinetics for these substrates. JNK1?1 followed a random sequential mechanism forming a ternary complex between JNK-substrate-ATP. K(m) for ATF2 and c-Jun was 1.1 and 2.8 ?M, respectively. Inhibition studies using adenosine 5'-(?,?-methylenetriphosphate) and a peptide derived from JNK interacting protein 1 (JIP1) supported the proposed kinetic mechanism. Biolayer interferometry studies showed that unphosphorylated JNK1?1 bound to ATF2 with similar affinity as it did to c-Jun (K(D) = 2.60 ± 0.34 versus 1.00 ± 0.35 ?M, respectively). The presence of ATP increased the affinity of unphosphorylated JNK1?1 for ATF2 and c-Jun, to 0.80 ± 0.04 versus 0.65 ± 0.07 ?M, respectively. Phosphorylation of JNK1?1 decreased the affinity of the kinase for ATF2 to 11.0 ± 1.1 ?M and for c-Jun to 17.0 ± 7.5 ?M in the absence of ATP. The presence of ATP caused a shift in the K(D) of the active kinase for ATF2 to 1.70 ± 0.25 ?M and for c-Jun of 3.50 ± 0.95 ?M. These results are the first kinetic and biochemical characterization of JNK1?1 and uncover some of the differences in the enzymatic activity of JNK1?1 compared with other variants and suggest that ATP binding or JNK phosphorylation could induce changes in the interactions with substrates, activators, and regulatory proteins.

SUBMITTER: Figuera-Losada M

PROVIDER: S-EPMC3339953 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature

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Enzyme kinetics and interaction studies for human JNK1β1 and substrates activating transcription factor 2 (ATF2) and c-Jun N-terminal kinase (c-Jun).

Figuera-Losada Mariana M LoGrasso Philip V PV

The Journal of biological chemistry 20120217 16

c-Jun N-terminal kinase (JNK) is a stress signal transducer linked to cell death, and survival. JNK1 has been implicated in obesity, glucose intolerance, and insulin resistance. In this study we report the kinetic mechanism for JNK1β1 with transcription factors ATF2 and c-Jun along with interaction kinetics for these substrates. JNK1β1 followed a random sequential mechanism forming a ternary complex between JNK-substrate-ATP. K(m) for ATF2 and c-Jun was 1.1 and 2.8 μM, respectively. Inhibition s ...[more]

PMID: 22351776

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Dataset Information

Enzyme kinetics and interaction studies for human JNK1?1 and substrates activating transcription factor 2 (ATF2) and c-Jun N-terminal kinase (c-Jun).

Publications

Enzyme kinetics and interaction studies for human JNK1β1 and substrates activating transcription factor 2 (ATF2) and c-Jun N-terminal kinase (c-Jun).

Similar Datasets

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