Initial purification study of the cytochromeb 561 of bean hypocotyl plasma membrane

Autor: Han Asard, I. Bäurle, Paolo Trost, L. Rotino, Sandra Scagliarini, Paolo Pupillo
Rok vydání: 1998
Předmět:
Zdroj: Protoplasma. 205:66-73
ISSN: 1615-6102
0033-183X
DOI: 10.1007/bf01279295
Popis: The plasma membrane (PM) of higher plants contains a major ascorbate-reducible, high-potentialb-type cytochrome, named cytochromeb561 (cytb561). In this paper a rapid purification protocol for the cytb561 of bean hypocotyls PM is described. An almost 200-fold increase of cytb561 specific concentration was achieved with respect to the PM fraction, which contained about 0.2 nmol of ascorbate-reducible heme per mg protein. The procedure can be performed in one day starting from purified PMs obtained by the phase-partitioning procedure. However, cytb561 proved to be unstable during chromatographic purification and the amount of protein finally recovered was low. Purified cytb561 eluted as a 130,000 Da protein-detergent complex from gel-filtration columns. It was completely reduced by ascorbate and reduced-minus-oxidized spectra showed α-, β- and γ-bands at 561, 530, and 429 nm respectively, not unlike the spectra of whole PMs. This work represents an initial approach to the biochemical characterization of the cytb561 of higher plants, formerly suggested to be related to cytb561 of animal chromaffin granules.
Databáze: OpenAIRE
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