Evidence for ligand-induced conformational changes in proteins from phosphorescence spectroscopy.
Phosphorescence spectroscopy on mouse myeloma IgA J539 in rigid solution at 77K revealed the type of anomalous short-lived component in the tryptophan decay originally observed with lysozyme (Churchich, J.E., 1966. Biochim. Biophys. Acta. 120:406-412) and seen in a large number of Bence Jones proteins (Longworth, J.W., C.L. McLaughlin, and A. Solomon. 1976. Biochemistry. 15:2953-2958). The decay time of the anomalous component that results from the interaction between tryptophan side chains and disulfide linkages in proteins was observed to significantly lengthen in J539 in response to binding of a galactan antigen. With hen egg-white lysozyme in which the type of fluorescence enhancement on ligand binding seen with J539 has also been observed, phosphorescence measurements revealed a similar lengthening of the decay time of the disulfide-induced anomalous component in the tryptophan decay. These perturbations are interpreted as ligand-induced changes to the tryptophan-disulfide proximities that have been shown to exist in these structures. Given the short-range nature of the disulfide perturbation (see following article) the observations suggest, in particular when combined with x-ray crystallographic data, that phosphorescence decay-time measurements of disulfide perturbations can serve as a sensitive spectroscopic indicator of subtle conformational changes in immunoglobulins and other tryptophan-disulfide containing proteins.
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Department of Chemistry, McGill University, Montreal, Quebec, Canada.
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D000936:Antigen-Antibody Complex; D004220:Disulfides; D007070:Immunoglobulin A; D008024:Ligands; D049449:Luminescence; D008958:Models, Molecular; D009113:Muramidase / Q000378:metabolism; D009194:Myeloma Proteins / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods; D014364:Tryptophan
D000936:Antigen-Antibody Complex; D004220:Disulfides; D007070:Immunoglobulin A; D008024:Ligands; D009194:Myeloma Proteins; D011506:Proteins; D014364:Tryptophan; D009113:Muramidase