Some peculiarities of the dynamics of the immunoglobulin M structure.
The isotropic mobility of separate regions of the intact molecule of immunoglobulin M (IgM) and its Fab and (Fc)5 fragments was studied using spin-labeling of carbohydrate (2,2,6,6-tetramethyl-4-aminopiperidine-1-oxyl) and peptide (2,2,5,5-tetramethyl-3-dichlorotriazinylaminopyrrolidine-1-oxyl) moieties. The spin-labeled oligosaccharide groups (OGs) in the Fab region are shown to have much more amplitude of anisotropic motion than those in the (Fc)5 region. The spin label in the latter is evidently attached in the C mu 3 domain to one of its OGs which is probably stabilized by ionic contacts between terminal N-acetylneuraminic acid residue and the peptide moiety of the IgM molecule. When the amount of the glycosidase-cleaved carbohydrate does not exceed 10-15%, most OGs affected are of the Fab region. Upon profound splitting (greater than or equal to 50%) the OGs of the (Fc)5 region are also affected; that results evidently in loosening the ionic contacts between the shortened OGs and the peptide moiety of IgM, and consequently in increasing mobility of the former. The structure of the (Fc)5 region of IgM is labile; after detaching this moiety from the intact IgM molecule, its structure is stabilized, but one of its domains (C mu 3) becomes more mobile than it is in the intact IgM molecule; at the same time the amplitude of anisotropic motion of OG bound here is decreased. In the latter case, this decrease depends on the sequence of spin-labeling and fragmentation. The most probable cause of stabilization of the (Fc)5 fragment is the heating of IgM solution to 56 degree C during fragmentation with trypsin. At this temperature the tau value for the (Fc)5 fragment is unusually low, equaling 23 ns. The spin-labeling in the peptide part of IgM occurs mostly in the Fab region which is a rather rigid moiety as expected.
D004578:Electron Spin Resonance Spectroscopy; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007075:Immunoglobulin M; D015394:Molecular Structure; D009844:Oligosaccharides; D011487:Protein Conformation
D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007075:Immunoglobulin M; D009844:Oligosaccharides