title: Effect of genetic variation on the tryptic hydrolysis of bovine beta-lactoglobulin A, B, and C.
abstract: The structure, stability, and hydrolysis characteristics of beta-lactoglobulin (LG) A are different from those of either beta-LG B or beta-LG C. Purified samples of these proteins were hydrolyzed with trypsin and the rates of loss of native monomeric beta-LG structure were measured using sodium dodecyl sulfate PAGE. At the same time, the appearance of many individual peptides were identified and followed in time by HPLC, measuring their concentration as a function of solution pH, temperature, protein concentration, and added urea or palmitate. The identity of the peptides was confirmed by liquid chromatography-mass spectrometry. This semiquantitative exploration showed that the rate of hydrolysis was in the order beta-LG A > beta-LG B > beta-LG C under most circumstances, and that 12 of the 18 trypsin-susceptible bonds were cleaved at very similar rates that were governed by the variant type. Consequently, the rate of hydrolysis of the intact protein was related to the overall structural stability of the individual proteins and the accessibility of certain peptide bonds to the enzyme. The hydrolysis of mixtures of 2 or more variants or of denatured beta-LG gave more heterogeneous peptide mixtures.
authors.affiliation: Fonterra Research Centre, P.O. Box 11 029, Palmerston North, New Zealand.
authors.forename: L K
authors.initials: LK
authors.lastname: Creamer
authors.forename: H C
authors.initials: HC
authors.lastname: Nilsson
authors.forename: M A
authors.initials: MA
authors.lastname: Paulsson
authors.forename: C J
authors.initials: CJ
authors.lastname: Coker
authors.forename: J P
authors.initials: JP
authors.lastname: Hill
authors.forename: R
authors.initials: R
authors.lastname: Jiménez-Flores
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D014644:Genetic Variation; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D007782:Lactoglobulins / Q000737:chemistry* / Q000235:genetics; D017433:Protein Structure, Secondary; D013696:Temperature; D014357:Trypsin / Q000737:chemistry*
chemical_list: D007782:Lactoglobulins; D014357:Trypsin