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Title: Purification and properties of glycoprotein processing. cap alpha. -mannosidase from mung bean seedlings

Abstract

The microsomal fraction of mung bean seedlings contains mannosidase activities capable of hydrolyzing (/sup 3/H) mannose from the (/sup 3/H)Man/sub 9/GlcNAc as well as for releasing mannose from p-nitrophenyl-..cap alpha..-D-mannopyranoside. The glycoprotein processing mannosidase was purified by conventional methods and also by affinity chromatography on mannan-Sepharose and mannosamine-Sepharose. The final enzyme preparation contained a trace of aryl-mannosidase, but this activity was inhibited by swainsonine whereas the processing enzyme was not. The pH optimum for the processing enzyme was 5.5 to 6.0, and activity was optimum in the presence of 0.1% Triton X-100. The enzyme was inhibited by ethylenediaminetetraacetate while Ca/sup 2 +/ was the most effective cation for reversing this inhibition. Mn/sup 2 +/ was considerably less effective than Ca/sup 2 +/ and Mg/sup 2 +/ was without effect. The processing mannosidase was inhibited by ..cap alpha..1,2- and ..cap alpha..1,3-linked mannose oligosaccharides whereas free mannose and ..cap alpha..1,6-linked mannose oligosaccharides were ineffective. Mannosamine was also an inhibitor of this enzyme. The aryl-mannosidase and the processing mannosidase could also be distinguished by their susceptibility to various processing inhibitors. The processing mannosidase was incubated for long periods with (/sup 3/H)Man/sub 9/GlcNAc and the products were identified by gel filtration. Even after amore » 24 hour incubation, the only two radioactive products were Man/sub 5/GlcNAc and free mannose. Thus, this enzyme appears to be similar to the animal processing enzyme, mannosidase I, and is apparently a specific ..cap alpha..1,2-mannosidase.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Univ. of Texas Health Science Center, San Antonio
OSTI Identifier:
6992606
Resource Type:
Journal Article
Journal Name:
Plant Physiol.; (United States)
Additional Journal Information:
Journal Volume: 81:2
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLYCOSYL HYDROLASES; CHEMICAL PROPERTIES; ENZYME ACTIVITY; PURIFICATION; ENZYME INHIBITORS; MUNGBEANS; SEEDLINGS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; BACTERIA; ENZYMES; FOOD; HYDROLASES; ISOTOPE APPLICATIONS; LABELLED COMPOUNDS; LEGUMINOSAE; MICROORGANISMS; PLANTS; RHIZOBIUM; SEEDS; VEGETABLES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Szumilo, T, Kaushal, G P, Hori, H, and Elbein, A D. Purification and properties of glycoprotein processing. cap alpha. -mannosidase from mung bean seedlings. United States: N. p., 1986. Web.
Szumilo, T, Kaushal, G P, Hori, H, & Elbein, A D. Purification and properties of glycoprotein processing. cap alpha. -mannosidase from mung bean seedlings. United States.
Szumilo, T, Kaushal, G P, Hori, H, and Elbein, A D. Sun . "Purification and properties of glycoprotein processing. cap alpha. -mannosidase from mung bean seedlings". United States.
@article{osti_6992606,
title = {Purification and properties of glycoprotein processing. cap alpha. -mannosidase from mung bean seedlings},
author = {Szumilo, T and Kaushal, G P and Hori, H and Elbein, A D},
abstractNote = {The microsomal fraction of mung bean seedlings contains mannosidase activities capable of hydrolyzing (/sup 3/H) mannose from the (/sup 3/H)Man/sub 9/GlcNAc as well as for releasing mannose from p-nitrophenyl-..cap alpha..-D-mannopyranoside. The glycoprotein processing mannosidase was purified by conventional methods and also by affinity chromatography on mannan-Sepharose and mannosamine-Sepharose. The final enzyme preparation contained a trace of aryl-mannosidase, but this activity was inhibited by swainsonine whereas the processing enzyme was not. The pH optimum for the processing enzyme was 5.5 to 6.0, and activity was optimum in the presence of 0.1% Triton X-100. The enzyme was inhibited by ethylenediaminetetraacetate while Ca/sup 2 +/ was the most effective cation for reversing this inhibition. Mn/sup 2 +/ was considerably less effective than Ca/sup 2 +/ and Mg/sup 2 +/ was without effect. The processing mannosidase was inhibited by ..cap alpha..1,2- and ..cap alpha..1,3-linked mannose oligosaccharides whereas free mannose and ..cap alpha..1,6-linked mannose oligosaccharides were ineffective. Mannosamine was also an inhibitor of this enzyme. The aryl-mannosidase and the processing mannosidase could also be distinguished by their susceptibility to various processing inhibitors. The processing mannosidase was incubated for long periods with (/sup 3/H)Man/sub 9/GlcNAc and the products were identified by gel filtration. Even after a 24 hour incubation, the only two radioactive products were Man/sub 5/GlcNAc and free mannose. Thus, this enzyme appears to be similar to the animal processing enzyme, mannosidase I, and is apparently a specific ..cap alpha..1,2-mannosidase.},
doi = {},
url = {https://www.osti.gov/biblio/6992606}, journal = {Plant Physiol.; (United States)},
number = ,
volume = 81:2,
place = {United States},
year = {1986},
month = {6}
}