Multinuclear NMR study of the structure of the Fv fragment of anti-dansyl mouse IgG2a antibody.
A multinuclear NMR study is reported of Fv, which is a minimum antigen-binding unit of immunoglobulin. Fv has been prepared by clostripain digestion of a mouse anti-dansyl IgG2a monoclonal antibody that lacks the entire CH1 domain [Takahashi, H., Igarashi, T., Shimada, I., & Arata, Y. (1991) Biochemistry 30, 2840-2847]. A variety of Fv analogues labeled with 2H in the aromatic rings and with 13C and/or 15N in the peptide bonds have been prepared and used for multinuclear NMR analyses of Fv in the absence and presence of epsilon-dansyl-L-lysine (DNS-Lys). It has been shown that 1H-15N shift correlation spectra of Fv sensitively reflect the antigen binding and can be used along with 1H and 13C spectral data for the structural analyses of antigen-antibody interactions. Hydrogen-deuterium exchange of the amide protons has been followed in the absence and presence of DNS-Lys by using the 1H-15N shift correlation spectra. Use of the beta-shift observed for the carbonyl carbon resonances has also been helpful in following the hydrogen-deuterium exchange. On the basis of the NMR data obtained, the static and dynamic structure of the Fv fragment in the absence and presence of DNS-Lys has been discussed.
Takahashi
H
H
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Odaka
A
A
Kawaminami
S
S
Matsunaga
C
C
Kato
K
K
Shimada
I
I
Arata
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001666:Binding Sites, Antibody; D003619:Dansyl Compounds; D004151:Dipeptides / Q000032:analysis; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G / Q000737:chemistry*; D007136:Immunoglobulins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry*; D014443:Tyrosine
D000911:Antibodies, Monoclonal; D003619:Dansyl Compounds; D004151:Dipeptides; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007136:Immunoglobulins; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D014443:Tyrosine