Folding-related dimerization of human cystatin C.
With the aim to improve our understanding of the structural basis for protein self-association and aggregation, in particular in relationship to protein refolding and amyloid formation, folding-related processes for human cystatin C have been studied. Using NMR spectroscopy together with chromatographic and electrophoretic methods, a self-association process resulting in dimer formation for protein samples treated with denaturing agents as well as for samples subjected to low pH or high temperature conditions could be studied with amino acid resolution. In all three cases, the dimerization involves properly folded molecules and proceeds via the reactive site of the inhibitor, which leads to complete loss of its biological activity. This dimerization process has potential relevance for amyloid formation by the brain hemorrhage-causing Leu58-Gln variant of cystatin C. The results also indicate that cystatin C dimerization and inactivation may occur in acidified compartments in vivo, which could be relevant for the physiological regulation of cysteine proteinase activity.
Ekiel
I
I
Pharmaceutical Biotechnology Sector, National Research Council of Canada, Montreal, Quebec, Canada.
Abrahamson
M
M
D000595:Amino Acid Sequence; D000682:Amyloid / Q000096:biosynthesis; D002543:Cerebral Hemorrhage / Q000235:genetics; D003001:Cloning, Molecular; D055316:Cystatin C; D015891:Cystatins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli; D014644:Genetic Variation; D005973:Glutamine; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007930:Leucine; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013816:Thermodynamics
D000682:Amyloid; C525030:CST3 protein, human; D055316:Cystatin C; D015891:Cystatins; D006146:Guanidines; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D005973:Glutamine; D007930:Leucine; D019791:Guanidine