Heat-induced redistribution of disulfide bonds in milk proteins. 1. Bovine beta-lactoglobulin.
Changes in the structure and chemistry of beta-lactoglobulin (beta-LG) play an important role in the processing and functionality of milk products. In model beta-LG systems, there is evidence that the aggregates of heated beta-LG are held together by a mixture of intermolecular non-covalent association and heat-induced non-native disulfide bonds. Although a number of non-native disulfide bonds have been identified, little is known about the initial inter- and intramolecular disulfide bond rearrangements that occur as a result of heating. These interchange reactions were explored by examining the products of heat treatment to determine the novel disulfide bonds that form in the heated beta-LG aggregates. The native protein and heat-induced aggregates were hydrolyzed by trypsin, and the resulting peptides, before and after reduction with dithiothreitol, were separated by high-performance liquid chromatography and their identities confirmed by electrospray ionization mass spectrometry. Comparisons of these peptide patterns showed that some of the Cys160 was in the reduced form in heated beta-LG aggregates, indicating that the Cys160-Cys66 disulfide bond had been broken during heating. This finding suggests that disulfide bond interchange reactions between beta-LG non-native monomers, or polymers, and other proteins could occur largely via Cys160.
Creamer
Lawrence K
LK
Fonterra Research Centre, Private Bag 11-029, Palmerston North, New Zealand. lawrie.creamer@fonterra.com
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Annie
A
Nilsson
Hanna
H
Paulsson
Marie
M
van Wanroij
Miriam
M
Lowe
Edwin K
EK
Anema
Skelte G
SG
Boland
Michael J
MJ
Jiménez-Flores
Rafael
R
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry*; D006358:Hot Temperature; D006868:Hydrolysis; D007782:Lactoglobulins / Q000737:chemistry* / Q000378:metabolism; D008894:Milk Proteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D021241:Spectrometry, Mass, Electrospray Ionization; D014357:Trypsin / Q000378:metabolism
D004220:Disulfides; D007782:Lactoglobulins; D008894:Milk Proteins; D010446:Peptide Fragments; D014357:Trypsin