Characterization of covalent protein conjugates using solid-state 13C NMR spectroscopy.
Cross-polarization magic-angle spinning (CPMAS) 13C NMR spectroscopy has been used to characterize covalent conjugates of alachlor, an alpha-chloroacetamide hapten, with glutathione (GSH) and bovine serum albumin (BSA). The solid-state NMR method demonstrates definitively the covalent nature of these conjugates and can also be used to characterize the sites of hapten attachment to proteins. Three different sites of alachlor binding are observed in the BSA system. Accurate quantitation of the amount of hapten covalently bound to GSH and BSA is reported. The solid-state 13C NMR technique can easily be generalized to study other small molecule/protein conjugates and can be used to assist the development and refinement of synthetic methods needed for the successful formation of such protein alkylation products.
Garbow
J R
JR
Monsanto Company, Life Sciences Research Center, St. Louis, Missouri 63198.
Fujiwara
H
H
Sharp
C R
CR
Logusch
E W
EW
D000081:Acetamides / Q000737:chemistry*; D002247:Carbon Isotopes; D003460:Crystallization; D005978:Glutathione / Q000737:chemistry*; D006241:Haptens / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D011107:Polylysine / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D000081:Acetamides; D002247:Carbon Isotopes; D006241:Haptens; C000188:alachlor; D011107:Polylysine; D012710:Serum Albumin, Bovine; D005978:Glutathione