Chemical modification of rabbit IgG with N-dansylaziridine. Investigation of the properties of dansylated antibodies.
To elucidate the effect of antigen binding fluorescent thiol reagent, N-dansylaziridine (DAZ), which is sensitive to the changes in the microenvironment, was used for modification of rabbit IgG hinge region cysteine residue. DAZ binds to hinge region Cys 226 as evidenced by the structural analysis. Labelling of IgG with DAZ does not alter its conformation, hydrodynamic behavior, nor its antigen binding properties. Upon antigen bindings, the fluorescence intensity of modified IgG increases about 80%. This result indicates that interaction of antibodies with antigen is accompanied by the conformational changes in the IgG hinge region.
Pikuleva
I A
IA
Institute of Bioorganic Chemistry, BSSR, Academy of Sciences, Minsk U.S.S.R.
Turko
I V
IV
Adamovich
T B
TB
Chashchin
V L
VL
D000595:Amino Acid Sequence; D000818:Animals; D000937:Antigen-Antibody Reactions; D001388:Aziridines / Q000737:chemistry; D002942:Circular Dichroism; D003619:Dansyl Compounds / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010434:Pepsin A / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds; D013954:Thyroglobulin / Q000276:immunology; D014357:Trypsin / Q000737:chemistry
D001388:Aziridines; D003619:Dansyl Compounds; D007074:Immunoglobulin G; D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; C017173:dansylaziridine; D013954:Thyroglobulin; D014357:Trypsin; D010434:Pepsin A