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The sulfhydryl protease B was isolated from the cotyledons of 8-day old vetch seedlings and purified 1580-fold with a 38% recovery. The preparation obtained proved to be homogeneous by DEAE-cellulose chromatography and polyacrylamide gel electrophoresis. The molecular weight of the enzyme as shown
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Protease A is 870-fold purified by means of isoelectric precipitation, DEAE-cellulose chromatography and gel filtration through Sephadex G-50, the yield of the enzyme being 28%. The purified preparation is free of contaminant proteolytic activity and is almost homogenous chromatographically, but it
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A cDNA clone encoding a thiol-protease (TPE4A) was isolated from senescent ovaries of pea (Pisum sativum) by reverse transcriptase-polymerase chain reaction. The deduced amino acid sequence of TPE4A has the conserved catalytic amino acids of papain. It is very similar to VSCYSPROA, a thiol-protease
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L-phenylalanine-p-nitroanilidase (PPA-ase) from vetch cotyledons was purified 1600-fold with a 6.7% recovery. Data from gel electrophoresis suggest that the preparation obtained (specific activity 232 U/mg) contains only a small admixture of inactive protein. PPA-ase splits off more than 14% of
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The split-off 1-2 short peptides is the first stop in the endogenous protease A effect on the vetch legumin, which results in a step-wise rise of its hydrolyzability by two other endogenous proteases (B and C). Short neutral and basic peptides are consecutively split off from the acid subunits in
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In vitro binding experiments were carried out using (32)P-labeled cells of the virulent Agrobacterium tumefaciens strain B6 and Datura innoxia cells from suspension culture. Binding kinetics showed that adherence of bacteria to Datura cells increased gradually during the first 60 minutes and
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NodO is a secreted protein from Rhizobium leguminosarum bv. viciae with a role in signalling during legume nodulation. A Tn5-induced mutant was identified that was defective in NodO secretion. As predicted, the secretion defect decreased pea and vetch nodulation but only when the nodE gene was also
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