Chloroplast biogenesis 84: solubilization and partial purification of membrane-bound [4-vinyl]chlorophyllide a reductase from etiolated barley leaves.
[4-Vinyl] chlorophyllide a reductase (4VCR) is a key enzyme of the chlorophyll (Chl) biosynthetic pathway. It catalyzes the conversion of divinyl chlorophyllide (Chlide) a to monovinyl Chlide a by reduction of the vinyl group at position 4 of the macrocycle to ethyl. 4VCR is a membrane-bound enzyme, embedded in etioplast and etiochloroplast membranes. A study of the regulation and properties of this enzyme is mandatory for a comprehensive understanding of the biosynthetic heterogeneity of Chl biosynthesis. Solubilization and partial purification of 4VCR are described for the first time. The enzyme was solubilized with 5 mM Chaps and was partially purified by chromatography on DEAE-Sephacel and Cibacron Blue 3GA-1000 agarose. An overall 20-fold purification was achieved. The partially purified enzyme was stable for several months at -80 degrees C.
Kolossov
V L
VL
Laboratory of Plant Biochemistry and Photobiology, NRES, 240 A ERML, University of Illinois, 1201 West Gregory Avenue, Urbana, IL 61801, USA.
Rebeiz
C A
CA
D002462:Cell Membrane / Q000201:enzymology; D002734:Chlorophyll / Q000096:biosynthesis; D002793:Cholic Acids; D002845:Chromatography / Q000379:methods; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005990:Glycerol; D001467:Hordeum; D008027:Light; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D018515:Plant Leaves / Q000201:enzymology; D012995:Solubility
D002793:Cholic Acids; D002734:Chlorophyll; D010088:Oxidoreductases; C097964:(4-vinyl)chlorophyllide a reductase; D005990:Glycerol; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; D004229:Dithiothreitol