Interaction of the heart-specific LIM domain protein, FHL2, with DNA-Binding nuclear protein, hNP220
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摘要Using a yeast two-hybrid library screen, we have identified that the heart specific FHL2 protein, four-and-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. (C) 2001 Wiley-Liss, Inc.
著者Ng EKO, Chan KK, Wong CH, Tsui SKW, Ngai SM, Lee SMY, Kotaka M, Lee CY, Waye MMY, Fung KP
期刊名稱Journal of Cellular Biochemistry
出版年份2002
月份1
日期1
卷號84
期次3
出版社WILEY-LISS
頁次556 - 566
國際標準期刊號0730-2312
電子國際標準期刊號1097-4644
語言英式英語
關鍵詞human heart cDNA; molecular adapter; protein-protein interaction; yeast two-hybrid system; zinc finger protein
Web of Science 學科類別Biochemistry & Molecular Biology; BIOCHEMISTRY & MOLECULAR BIOLOGY; Cell Biology; CELL BIOLOGY

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