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Interaction of the heart-specific LIM domain protein, FHL2, with DNA-binding nuclear protein, hNP220
Ng E.K.O.; Chan K.K.; Wong C.H.; Tsui S.K.W.; Ngai S.M.; Lee S.M.Y.; Kotaka M.; Lee C.Y.; Waye M.M.Y.; Fung K.P.
2001-12-01
Source PublicationJournal of Cellular Biochemistry
ISSN07302312
Volume84Issue:3Pages:556-566
Abstract

Using a yeast two-hybrid library screen, we have identified that the heart specific FHL2 protein, fourand-a-half LIM protein 2, interacted with human DNA-binding nuclear protein, hNP220. Domain studies by the yeast two-hybrid interaction assay revealed that the second LIM domain together with the third and the fourth LIM domains of FHL2 were responsible to the binding with hNP220. Using green fluorescent protein (GFP)-FHL2 and blue fluorescent protein (BFP)-hNP220 fusion proteins co-expressed in the same cell, we demonstrated a direct interaction between FHL2 and hNP220 in individual nucleus by two-fusion Fluorescence Resonance Energy Transfer (FRET) assay. Besides, Western blot analysis using affinity-purified anti-FHL2 antipeptide antibodies confirmed a 32-kDa protein of FHL2 in heart only. Virtually no expression of FHL2 protein was detected in brain, liver, lung, kidney, testis, skeletal muscle, and spleen. Moreover, the expression of FHL2 protein was also detectable in the human diseased heart tissues. Our results imply that FHL2 protein can shuttle between cytoplasm and nucleus and may act as a molecular adapter to form a multicomplex with hNP220 in the nucleus, thus we speculate that FHL2 may be particularly important for heart muscle differentiation and the maintenance of the heart phenotype. © 2001 Wiley-Liss, Inc.

KeywordHuman Heart Cdna Molecular Adapter Protein-protein Interaction Yeast Two-hybrid System Zinc Finger Protein
DOI10.1002/jcb.10041
URLView the original
Indexed BySCI
WOS Research AreaBiochemistry & Molecular Biology ; Cell Biology
WOS SubjectBiochemistry & Molecular Biology ; Cell Biology
WOS IDWOS:000173358300012
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Cited Times [WOS]:27   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
AffiliationChinese University of Hong Kong
Recommended Citation
GB/T 7714
Ng E.K.O.,Chan K.K.,Wong C.H.,et al. Interaction of the heart-specific LIM domain protein, FHL2, with DNA-binding nuclear protein, hNP220[J]. Journal of Cellular Biochemistry,2001,84(3):556-566.
APA Ng E.K.O..,Chan K.K..,Wong C.H..,Tsui S.K.W..,Ngai S.M..,...&Fung K.P..(2001).Interaction of the heart-specific LIM domain protein, FHL2, with DNA-binding nuclear protein, hNP220.Journal of Cellular Biochemistry,84(3),556-566.
MLA Ng E.K.O.,et al."Interaction of the heart-specific LIM domain protein, FHL2, with DNA-binding nuclear protein, hNP220".Journal of Cellular Biochemistry 84.3(2001):556-566.
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