Purification of chloroplast alpha-1,4-glucan phosphorylase from spinach leaves by chromatography on Sepharose-bound starch.
Biochim Biophys Acta
; 659(1): 123-31, 1981 May 14.
Article
em En
| MEDLINE
| ID: mdl-7248312
ABSTRACT
Chloroplast alpha-1,4-glucan phosphorylase (EC 2.4.1.1) has been purified to homogeneity from spinach leaves as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purification procedure is composed of (NH4)2SO4 precipitation, ion-exchange chromatography, and chromatography on Sepharose-bound starch. In order to achieve binding of the chloroplast phosphorylase, a previously described Sepharose-glucan gel (Steup, M. Schächtele, C. and Latzko, E. (1980) Planta 148, 168-173) was modified by introducing hydrophobic groups in addition to the covalently bound starch. The chloroplast phosphorylase exhibited complete binding to this type of gel and could be eluted by a mixture of soluble glucan and NaCl. For the purified chloroplast phosphorylase, sodium dodecyl-sulfate polyacrylamide gel electrophoresis and pyridoxal phosphate determination resulted in a molecular weight estimation of about 110,000 per monomer. The apparent molecular weight of the native enzyme, as determined by polyacrylamide density gradient electrophoresis and gel filtration on Sephadex G-200, was 200,000 and 220,000, respectively. The data indicate that the chloroplast phosphorylase is a dimer with a molecular weight higher than that of the non-chloroplast phosphorylase.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Plantas
/
Fosforilases
Idioma:
En
Ano de publicação:
1981
Tipo de documento:
Article