Residues Cys-1 and Cys-79 are not essential for refolding of reduced-denatured kringle 4 fragment of human plasminogen.
It was shown previously that the Cys-1-Cys-79 disulphide bond forms in the last step of refolding of kringle 4 and that this bond is not essential for the lysine-Sepharose affinity of the kringle 4 fragment (Trexler, M. and Patthy, L. (1983) Proc. Natl. Acad. Sci. U.S.A. 80 2457-2461). Here we show that kringle 4, carboxymethylated on Cys-1 and Cys-79, regains its lysine-Sepharose affinity following denaturation and reductive cleavage of its disulphide bonds. The rate of refolding under aerobic conditions or in the presence of oxidized and reduced glutathione was similar to that observed in the case of native kringle 4. Our results suggest that Cys-1 and Cys-79 residues of kringles are not essential for the maintenance or acquisition of the biologically active kringle-fold.
D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D003553:Cystine / Q000032:analysis*; D004226:Dithioerythritol / Q000494:pharmacology; D006801:Humans; D007525:Isoelectric Focusing; D010446:Peptide Fragments / Q000032:analysis*; D010958:Plasminogen / Q000032:analysis*; D011487:Protein Conformation; D011489:Protein Denaturation; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D010446:Peptide Fragments; D003553:Cystine; D004226:Dithioerythritol; D010958:Plasminogen; D014357:Trypsin