Co-localization of glyceraldehyde-3-phosphate dehydrogenase with ferredoxin-NADP reductase in pea leaf chloroplasts.
In immunogold double-labeling of pea leaf thin sections with antibodies raised against ferredoxin-NADP reductase (EC 1.18.1.2, FNR) and antibodies directed against the A or B subunits of the NADP-linked glyceraldehyde-3-P dehydrogenase (GAPD) (EC 1.2.1.13), many small and large gold particles were found together over the chloroplasts. Nearest neighbor analysis of the distribution of the gold particles indicates that FNR and the NADP-linked GAPD are co-localized, in situ. This suggests that FNR might carry FADH2 or NADPH from the thylakoid membrane to GAPD, or that ferredoxin might carry electrons to FNR co-localized with GAPD in the stroma. Crystal structures of the spinach enzymes are available. When they are docked computationally, the proteins appear, as modeled, to be able to form at least two different complexes. One involves a single GAPD monomer and an FNR monomer (or dimer). The amino acid residues located at the putative interface are highly conserved on the chloroplastic forms of both enzymes. The other potential complex involves the GAPD A2B2 tetramer and an FNR monomer (or dimer). The interface residues are conserved in this model as well. Ferredoxin is able to interact with FNR in either complex.
Negi
Surendra S
SS
Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Carol
Andrew A
AA
Pandya
Shivangi
S
Braun
Werner
W
Anderson
Louise E
LE
D000595:Amino Acid Sequence; D002736:Chloroplasts / Q000201:enzymology* / Q000648:ultrastructure; D003198:Computer Simulation; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D008854:Microscopy, Electron; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology*; D018515:Plant Leaves / Q000201:enzymology*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D005287:Ferredoxin-NADP Reductase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases