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[35] Use of cross-linking reagents to study lipoprotein structure
Book chapter   Peer reviewed

[35] Use of cross-linking reagents to study lipoprotein structure

John B. Swaney
Methods in Enzymology, pp 613-626
1986
PMID: 3724526

Abstract

A wide variety of cross-linking reagents have been utilized for studies of protein structure, and the introduction of covalent bonds between protein chains through the use of bifunctional reagents is useful for analysis of the quaternary structure of complex enzymes. This chapter describes cross-linking reagents, and the cross-linking methodology used for the study of apolipoprotein self-association and recombinant lipoprotein structure. The majority of cross-linking studies have been performed on intact lipoproteins and on apolipoproteins that have utilized bifunctional reagents with imidoesters as the active functional groups, for example, dimethyl suberimidate (DMS); however, the reagent of choice for analysis of nearest-neighbor proteins in lipoproteins is difluorodinitrobenzene (DFDNB). The cross-linking reaction is monitored by performing SDS gel electrophoresis and quantitating the extent of oligomer formation by scanning stained gels. There are a substantial number of areas in which chemical cross-linking can be expected to add useful insights, and one of these is in elucidating the structure of the larger lipoprotein classes; for example, cross-linking of very low-density lipoprotein might indicate the topographical proximity of the numerous protein components to one another.

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Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
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