Detergents as probes of hydrophobic binding cavities in serum albumin and other water-soluble proteins.
As an extension of our studies on the interaction of detergents with membranes and membrane proteins, we have investigated their binding to water-soluble proteins. Anionic aliphatic compounds (dodecanoate and dodecylsulfate) were bound to serum albumin with high affinity at nine sites; related nonionic detergents (C12E8 and dodecylmaltoside) were bound at seven to eight sites, many in common with those of dodecanoate. The compounds were also bound in the hydrophobic cavity of beta-lactoglobulin, but not to ovalbumin. In addition to the generally recognized role of the Sudlow binding region II of serum albumin (localized at the IIIA subdomain) in fatty acid binding, quenching of the fluorescence intensity of tryptophan-214 by 7,8-dibromododecylmaltoside and 12-bromododecanoate also implicate the Sudlow binding region I (subdomain IIA) as a locus for binding of aliphatic compounds. Our data document the usefulness of dodecyl amphipathic compounds as probes of hydrophobic cavities in water-soluble proteins. In conjunction with recent x-ray diffraction analyses of fatty acid binding as the starting point we propose a new symmetrical binding model for the location of nine high-affinity sites on serum albumin for aliphatic compounds.
Kragh-Hansen
U
U
Department of Medical Biochemistry, University of Aarhus, DK-8000 Aarhus C, Denmark.
Hellec
F
F
de Foresta
B
B
le Maire
M
M
Møller
J V
JV
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002645:Chickens; D003902:Detergents / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D007782:Lactoglobulins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D015335:Molecular Probes / Q000378:metabolism*; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D016415:Sequence Alignment; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism*; D012995:Solubility; D055672:Static Electricity; D013816:Thermodynamics; D014364:Tryptophan / Q000378:metabolism; D014867:Water / Q000378:metabolism*
D003902:Detergents; D005227:Fatty Acids; D007782:Lactoglobulins; D008024:Ligands; D015335:Molecular Probes; D012709:Serum Albumin; D014867:Water; D014364:Tryptophan; D010047:Ovalbumin