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Effect of genetic variation on the tryptic hydrolysis of bovine beta-lactoglobulin A, B, and C

Creamer, L K; Nilsson, H C; Paulsson, Marie LU ; Coker, C J; Hill, J P and Jimenez-Flores, R (2004) In Journal of Dairy Science 87(12). p.4023-4032
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
ß-lactoglobulin, tryptic hydrolysis, genetic variant, kinetics
in
Journal of Dairy Science
volume
87
issue
12
pages
4023 - 4032
publisher
American Dairy Science Association
external identifiers
  • wos:000225227800004
  • pmid:15545362
ISSN
1525-3198
language
English
LU publication?
yes
id
db9dab46-33bd-45b8-8196-9e6c72ff171c (old id 139151)
alternative location
http://jds.fass.org/cgi/content/abstract/87/12/4023
date added to LUP
2007-07-18 13:33:36
date last changed
2016-04-15 19:53:46
@article{db9dab46-33bd-45b8-8196-9e6c72ff171c,
  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.},
  author       = {Creamer, L K and Nilsson, H C and Paulsson, Marie and Coker, C J and Hill, J P and Jimenez-Flores, R},
  issn         = {1525-3198},
  keyword      = {ß-lactoglobulin,tryptic hydrolysis,genetic variant,kinetics},
  language     = {eng},
  number       = {12},
  pages        = {4023--4032},
  publisher    = {American Dairy Science Association},
  series       = {Journal of Dairy Science},
  title        = {Effect of genetic variation on the tryptic hydrolysis of bovine beta-lactoglobulin A, B, and C},
  volume       = {87},
  year         = {2004},
}