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The interaction of nelin, a cardiac-specific expressed protein of human novel gene nelin, with F-actin was studied by both F-actin cosedimentation in vitro and colocal-ization assays.The results showed that nelin is a new F-actin binding protein and is colocolized with F-actin in cytoplasm of cells.Three new nelin binding proteins, filamin C subtype, titin N2B subtype and inter-alpha trypsin inhibitor heavy chain precursor (ITIH), were identified from human heart cDNA library using yeast two-hybrid screening.The binding activity of filamin C with nelin was confirmed by coim-munoprecipitation.Filamin C binds to nelin through its C-terminal region.It is indicated that nelin is a cytoskeleton associated protein and acts as a membrane-cytoskeleton as-sociated protein involved in the formation of focal adhesions.