Open AccessPurification and Developmental Analysis of a Metalloendoproteinase from the Leaves of Glycine max
Author(s) -
John S. Graham,
Jin Xiong,
Jeffery W. Gillikin
Publication year - 1991
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.97.2.786
Subject(s) - glycine , size exclusion chromatography , electrophoresis , molecular mass , enzyme , biochemistry , chromatography , chemistry , gel electrophoresis , polyacrylamide gel electrophoresis , extracellular , chelation , biology , amino acid , organic chemistry
A metalloendoproteinase from leaves of soybean (Glycine max) has been purified 1160-fold to electrophoretic homogeneity. The native protein is monomeric with a molecular mass of 15 kilodaltons as estimated by gel filtration and 19 kilodaltons as estimated by denaturing gel electrophoresis. The enzyme has a pH optima of 8.0 to 9.0 using Azocoll as substrate. The proteolytic activity is susceptible to metal chelating agents and the inactivated enzyme can be restored to 69% of original activity by the addition of ZnCl(2). Western analysis shows that a fraction of the soybean metalloendoproteinase is present within the extracellular space of older leaves. Soybean metalloendoproteinase 1 is the Azocollase A activity first described by Ragster and Chrispeels (Plant Physiol 64: 857-862; 1979).