Protein-Protein Interaction of Zinc Finger LIM Domains with Protein Kinase C

doi:10.1074/jbc.271.49.31029

Protein-Protein Interaction of Zinc Finger LIM Domains with Protein Kinase C*

From the Biosignal Research Center, Kobe University, Kobe 657, Japan the
^§Department of Biology, Faculty of Science, Kyushu University, Fukuoka 812-81, Japan, the
^¶Inheritance and Variation Group, PRESTO, Research Development Corporation of Japan, Kyoto 619-02, Japan, and the
^∥Department of Medicine, University of California San Diego, La Jolla, California 92093-0650

^† To whom correspondence should be addressed:
Biosignal Research Center, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657, Japan.
Tel.: 81-78-803-1255; Fax: 81-78-803-0994; E-mail: skuroda{at}inherit.biosig.kobe-u.ac.jp.

Abstract

The LIM domain comprising two zinc-finger motifs is found in a variety of proteins and has been proposed to direct protein-protein interactions. During the identification of protein kinase C (PKC)-interacting proteins by a yeast two-hybrid assay, a novel protein containing three LIM domains, designated ENH, was shown to associate with PKC in an isoform-specific manner. Deletion analysis demonstrated that any single LIM domain of ENH associates with the NH₂-terminal region of PKC. ENH associated with PKC in COS-7 cells and was phosphorylated by PKC in vitro. Upon treatment of the cells with phorbol ester, ENH in the membrane fraction was translocated to the cytosol fraction in vivo. Other LIM domain-containing proteins, such as Enigma and LIM-kinase 1, also interacted with PKC through their LIM domains. These results suggest that the LIM domain is one of the targets of PKC and that the LIM-PKC interaction may shed light on undefined roles of LIM domain-containing proteins.

Footnotes

↵* This study was supported in part by research grants from the Scientific Research Funds of the Ministry of Education, Science, Sports and Culture of Japan, the Charitable Trust Osaka Cancer Researcher Fund, and the Yamanouchi Foundation for Research on Metabolic Disorders. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

GDNF

glial-cell-line-derived neurotrophic factor

GDNFR

GDNF receptor

InsR

insulin receptor

PKC

protein kinase C

PAGE

polyacrylamide gel electrophoresis

kb

kilobases

TPA

12-O-tetradecanoylphorbol-13-acetate

LIMK1

LIM-kinase 1

GST

glutathione S-transferase.
- Received August 28, 1996.
- Revision received September 30, 1996.