N-methyl-N-(5-carboxypentyl)-1-deoxynojirimycin, a new affinity ligand for the purification of trimming glucosidase I.
The paper describes the synthesis of a new type of affinity resin containing N-methyl-N-(5-carboxypentyl)-1-deoxynojirimycin as the ligand attached to AH-Sepharose 4B, which allows the purification of trimming glucosidase I from a detergent extract of pig liver crude microsomes in one step and with high yield. The structure of the affinity ligand was designed on the basis of the observation that N,N-dialkylated derivatives of 1-deoxynojirimycin do strongly inhibit trimming glucosidase I, but not nonspecific aryl-alpha-glucosidases, including glucosidase II. The specific binding of glucosidase I eliminates the need of additional purification steps with their associated losses which were required with the previously synthesized N-5(-carboxypentyl)-AH-Sepharose 4B resin in order to achieve a homogenous enzyme preparation.
Bause
E
E
Institut für Physiologische Chemie der Universität, Bonn, Germany.
Gross
A
A
Schweden
J
J
D017485:1-Deoxynojirimycin; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000737:chemistry; D065089:Glycoside Hydrolase Inhibitors; D007700:Kinetics; D008024:Ligands; D008099:Liver / Q000201:enzymology; D013329:Structure-Activity Relationship; D013552:Swine; D000520:alpha-Glucosidases / Q000302:isolation & purification* / Q000378:metabolism
D065089:Glycoside Hydrolase Inhibitors; D008024:Ligands; D017485:1-Deoxynojirimycin; C042059:glucosidase I; D000520:alpha-Glucosidases; D005944:Glucosamine