Carbon-13 NMR study of switch variant anti-dansyl antibodies: antigen binding and domain-domain interactions.
A 13C NMR study is reported of switch variant anti-dansyl antibodies, which possess the identical VH, VL, and CL domains in conjunction with highly homologous but not identical heavy-chain constant regions. Each of these antibodies has been selectively labeled with 13C at the carbonyl carbon of Trp, Tyr, His, or Cys residue by growing hybridoma cells in serum-free medium. Spectral assignments have been made by following the procedure described previously for the switch variant antibodies labeled with [1-13C]Met [Kato, K., Matsunaga, C., Igarashi, T., Kim, H., Odaka, A., Shimada, I., & Arata, Y. (1991) Biochemistry 30, 270-278]. On the basis of the spectral data collected for the antibodies and their proteolytic fragments, we discuss how 13C NMR spectroscopy can be used for the structural analyses of antigen binding and also of domain-domain interactions in the antibody molecule.
Kato
K
K
Faculty of Pharmaceutical Sciences, University of Tokyo, Hongo, Japan.
Matsunaga
C
C
Odaka
A
A
Yamato
S
S
Takaha
W
W
Shimada
I
I
Arata
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001666:Binding Sites, Antibody; D002247:Carbon Isotopes; D002460:Cell Line; D003619:Dansyl Compounds; D014644:Genetic Variation; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism; D007134:Immunoglobulin Switch Region; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation
D002247:Carbon Isotopes; D003619:Dansyl Compounds; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G