Investigation of the cooperative structure of Fc fragments from myeloma immunoglobulin G.
The cooperative structure of Fc fragments prepared from myeloma human IgG1 was studied using scanning microcalorimetry and fluorescence at pH 4.2-8.0. It was shown that the first to be melted are CH2 domains whose interaction with each other is rather weak, while that with CH3 domains is strong. Then CH3 domains which form a single cooperative block are melted. The data for the structure of the Fc fragment in solution agree with the X-ray data according to which the interaction between CH2 domains is mediated by the carbohydrate moiety while the two CH3 domains are strongly associated. The presence of intensive CH2-CH3 interaction is a distinctive feature of the state of the Fc fragment in the given pH region as compared to that at pH <4.1 [Tischenko, V. M., et al. (1982) Eur. J. Biochem. 126, 517-521; Ryazantsev, S., et al. (1990) Eur. J. Biochem. 190, 393-399]. First, cis interactions greatly increase the free energy of the native structure stabilization in CH2 domains. Second, they decrease this energy for CH3 domains when compared to the state of the latter at pH 3.8 or within the Fc' fragment (the dimer of CH3 domains). The temperature and enthalpy of melting of CH2 domains coincide in all the samples studied despite heterogeneity of the carbohydrate moiety. Thus, it may be postulated that the conservative part of CH2 domains makes a cardinal contribution to the interaction of these domains with the carbohydrate moiety.
Tischenko
V M
VM
Institute of Immunology, Moscow Region, Russia. tischen@sun.ipr.serpukhov.su
Abramov
V M
VM
Zav'yalov
V P
VP
D002152:Calorimetry, Differential Scanning; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000096:biosynthesis; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology; D009194:Myeloma Proteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
D007106:Immune Sera; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D009194:Myeloma Proteins; D016650:Fluorescein-5-isothiocyanate