Electrospray ionization mass spectrometry as a tool for determination of drug binding sites to human serum albumin by noncovalent interaction.
Most proteins in blood plasma bind ligands. Human serum albumin (HSA) is the main transport protein with a very high capacity for binding of endogenous and exogenous compounds in plasma. Many pharmacokinetic properties of a drug depend on the level of binding to plasma proteins. This work reports studies of noncovalent interactions by means of nanoelectrospray ionization mass spectrometry (nanoESI-MS) for determination of the specific binding of selected drug candidates to HSA. Warfarin, iopanoic acid and digitoxin were chosen as site-specific probes that bind to the main sites of HSA. Two drug candidates and two known binders to HSA were analyzed using a competitive approach. The drugs were incubated with the target protein followed by addition of site-specific probes, one at a time. The drug candidates showed predominant affinity to site I (warfarin site). Naproxen and glyburide showed affinity to both sites I and II. The advantages of nanoESI-MS for these studies are the sensitivity, the absence of labeled molecules and the short method development time.
Benkestock
Kurt
K
Biovitrum AB, Department of Structural Chemistry, SE-112 76, Stockholm, Sweden. kurt.benkestock@medivir.se
Edlund
Per-Olof
PO
Roeraade
Johan
J
D001665:Binding Sites; D004074:Digitoxin / Q000378:metabolism; D005905:Glyburide / Q000378:metabolism; D006801:Humans; D007480:Iopanoic Acid / Q000378:metabolism; D009288:Naproxen / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014859:Warfarin / Q000378:metabolism
D004364:Pharmaceutical Preparations; D012709:Serum Albumin; D009288:Naproxen; D014859:Warfarin; D004074:Digitoxin; D007480:Iopanoic Acid; D005905:Glyburide