title: Chlorophyll Breakdown in Senescent Chloroplasts (Cleavage of Pheophorbide a in Two Enzymic Steps).
abstract: The cleavage of pheophorbide (Pheide) a into primary fluoescent chlorophyll (Chl) catabolites (pFCCs) in senescent chloroplasts was investigated. Chloroplast preparations isolated from senescent canola (Brassica napus) cotyledons exhibited light-dependent production of pFCC when assay mixtures were supplemented with ferredoxin (Fd). pFCC production in detergent-solubilized membranes was dependent on the presence of an Fd-reducing system. Pheide a cleavage required the action of two proteins, Pheide a oxygenase and a stroma protein. In the absence of stroma protein, Pheide a oxygenase converted Pheide a into a red Chl catabolite (RCC), the presumptive intermediary product of Pheide a cleavage. Incubation of the stroma protein (RCC reductase) together with chemically synthesized RCC resulted in the production of three different FCCs. Two of these catabolites were identical to the pFCCs from canola or barley (Hordeum vulgare) (pFCC-1) and sweet pepper (Capsicum annuum) (pFCC-2), respectively. Thus, the conversion of Pheide a to pFCC could be demonstrated to proceed in two consecutive steps, and both reactions depended on reduced Fd as the source of electrons. The function of Fd in Chl breakdown in vivo is corroborated by the marked retention of this protein until the late stages of senescence, as demonstrated by immunoblotting.
authors.affiliation: Department of Plant Biology, University of Zurich, CH-8008 Zurich, Switzerland (S.R., D.M., P.M., S.H.).
authors.forename: S.
authors.initials: S
authors.lastname: Rodoni
authors.forename: W.
authors.initials: W
authors.lastname: Muhlecker
authors.forename: M.
authors.initials: M
authors.lastname: Anderl
authors.forename: B.
authors.initials: B
authors.lastname: Krautler
authors.forename: D.
authors.initials: D
authors.lastname: Moser
authors.forename: H.
authors.initials: H
authors.lastname: Thomas
authors.forename: P.
authors.initials: P
authors.lastname: Matile
authors.forename: S.
authors.initials: S
authors.lastname: Hortensteiner