INTERSUBUNIT INTERACTIONS IN THE BOVINE MITOCHONDRIAL COMPLEX-I AS REVEALED BY LIGAND BLOTTING

Bovine mitochondrial complex I (NADH:ubiquinone oxidoreductase) is com posed of 3 structural domains, designated FP (flavoprotein, 3 subunits ), IP (iron-sulfur protein, 7-8 subunits) and HP (hydrophobic protein, > 30 subunits). IP intervenes between FP and HP, and in complex I its 75 kDa subunit appears to interact with the 51 kDa subunit of FP. In this study, we show by the technique of ligand blotting that isolated IP binds (a) only to the 51 kDa subunit of FP, and (b) to the 42, 39, 23, 20 and 16 kDa subunits of HP. Because a 23 kDa and a 20 kDa subuni t of complex I are potential iron-sulfur proteins: these and our previ ous results are consistent with the following possible path of electro ns in complex I: NADH --> 51 and 24 kDa subunit of FP --> 75 kDa subun it of IP --> 23 and 20 kDa subunits of HP --> ubiquinone. (C) 1996 Aca demic Press, Inc.