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. 2008 Dec 12;283(50):34753–34761. doi: 10.1074/jbc.M807323200

FIGURE 1.

FIGURE 1.

AcGFP1-tagging of CI subunits. A, current CI assembly model. Biogenesis of the hydrogenase module (containing NDUFS3) is thought to form the starting point of assembly (steps 1, 2, and 3). This module is coupled to a membrane module (at step 4), which contains at least ND1. Later in assembly, this membrane module is completed by other hydrophobic subunits (i.e. NDUFB6). Subunits of the NADH dehydrogenase module (containing NDUFV1, NDUFV2) are presumably added after formation of subcomplex 6. Accessory subunits (NDUFA2, NDUFA12) are also thought to enter in a late stage. B, schematic representation of the AcGFP1-tagged CI subunits (fusion proteins) expressed by inducible HEK293 cell lines. Colors correspond to the functional modules in A. C, BN-PAGE fluorograms (f) and anti-EGFP antibody immunodetection (a) of different cell lines expressing low levels of the AcGFP1-tagged CI subunits. The position of holo-CI is marked with closed arrowheads. Open arrowheads (NDUFS3 cell line) indicate CI assembly intermediates (1–6) or monomeric (AcGFP1-NDUFS3) subunit (m).