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J Biol Chem, Vol. 273, Issue 52, 35102-35108, December 25, 1998

Chlorophyll a Formation in the Chlorophyll b Reductase Reaction Requires Reduced Ferredoxin

From the Botanisches Institut der Ludwig-Maximilians-Universität München, Menzingerstraße 67, 80638 München, Germany

The reduction of chlorophyllide b and its analogue zinc pheophorbide b in etioplasts of barley (Hordeum vulgare L.) was investigated in detail. In intact etioplasts, the reduction proceeds to chlorophyllide a and zinc pheophorbide a or, if incubated together with phytyldiphosphate, to chlorophyll a and zinc pheophytin a, respectively. In lysed etioplasts supplied with NADPH, the reduction stops at the intermediate step of 7¹-OH-chlorophyll(ide) and Zn-7¹-OH-pheophorbide or Zn-7¹-OH-pheophytin. However, the final reduction is achieved when reduced ferredoxin is added to the lysed etioplasts, suggesting that ferredoxin is the natural cofactor for reduction of chlorophyll b to chlorophyll a. The reduction to chlorophyll a requires ATP in intact etioplasts but not in lysed etioplasts when reduced ferredoxin is supplied. The role of ATP and the significance of two cofactors for the two steps of reduction are discussed.

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