Interaction of hCLIM1, an enigma family protein, with alpha-actinin 2.
Enigma proteins are proteins that possess a PDZ domain at the amino terminal and one to three LIM domains at the carboxyl terminal. They are cytoplasmic proteins that are involved with the cytoskeleton and signal transduction pathway. By virtue of the two protein interacting domains, they are capable of protein-protein interactions. Here we report a study on a human Enigma protein hCLIM1, in particular. Our study describes the interaction of the human 36 kDa carboxyl terminal LIM domain protein (hCLIM1), the human homologue of CLP36 in rat, with alpha-actinin 2, the skeletal muscle isoform of alpha-actinin. hCLIM1 protein was shown to interact with alpha-actinin 2 by yeast two-hybrid screening and immunochemical analyses. Yeast two-hybrid analyses also demonstrated that the LIM domain of hCLIM1 binds to the EF-hand region of alpha-actinin 2, defining a new mode of LIM domain interactions. Immunofluorescent study demonstrates that hCLIM1 colocalizes with alpha-actinin at the Z-disks in human myocardium. Taken together, our experimental results suggest that hCLIM1is a novel cytoskeletal protein and may act as an adapter that brings other proteins to the cytoskeleton.
Kotaka
M
M
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.
Kostin
S
S
Ngai
S
S
Chan
K
K
Lau
Y
Y
Lee
S M
SM
Li
H y
Hy
Ng
E K
EK
Schaper
J
J
Tsui
S K
SK
Fung
K p
Kp
Lee
C y
Cy
Waye
M M
MM
D000185:Actinin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D060588:LIM Domain Proteins; D018613:Microscopy, Confocal; D009132:Muscles / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D051381:Rats; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000378:metabolism
C401401:ACTN2 protein, human; C498530:Actn2 protein, rat; C484572:LDB2 protein, human; D060588:LIM Domain Proteins; D020033:Protein Isoforms; D014157:Transcription Factors; D000185:Actinin