Structure-function relationship in glycosylated alpha-chymotrypsin as probed by IMAC and IMACE.
Chemical glycosylation of bovine alpha-chymotrypsin, by a glucosamine adduct on the carboxyl group, results in the modification of its catalytic activity. The structural alterations of alpha-chymotrypsin resulting from its glycosylation are studied by immobilized metal-ion affinity chromatography (IMAC) and immobilized metal-ion affinity capillary electrophoresis (IMACE). The chemical glycosylation of alpha-chymotrypsin generates two distinct subpopulations of the protein: one which totally loses the initial affinity for IDA-Cu(II) and another which exhibits an increased affinity for the metal chelate ligand.
Jiang
K Y
KY
Laboratoire d'Interactions Moléculaires et de Technologie des Séparations (LIMTech.S), Université de Technologie de Compiègne, CNRS-UPRES A 6022, Centre de Recherche de Royallieu BP 529, 60 205, Compiègne, Cedex, France.
Pitiot
O
O
Anissimova
M
M
Adenier
H
H
Vijayalakshmi
M A
MA
D000818:Animals; D002364:Caseins / Q000737:chemistry; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002918:Chymotrypsin / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005944:Glucosamine / Q000737:chemistry; D006031:Glycosylation; D006639:Histidine / Q000737:chemistry; D006860:Hydrogen Bonding; D007098:Imino Acids / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008670:Metals; D013329:Structure-Activity Relationship; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry
D002364:Caseins; D007098:Imino Acids; D008670:Metals; C030699:copper(II)-iminodiacetate; C006716:benzoyltyrosine ethyl ester; D014443:Tyrosine; D006639:Histidine; D002918:Chymotrypsin; D005944:Glucosamine