The role of histidyl residues in zinc-induced precipitation of alpha 2-macroglobulin-proteinase complexes.
When alpha 2-macroglobulin (alpha 2M) is reacted with proteinases including trypsin, plasmin, alpha-thrombin, or with CH3NH2, each resulting alpha 2M derivative is precipitated by Zn2+ in a similar manner. By contrast, unreacted alpha 2M is not precipitated over the same Zn2+ concentration range. Zn2+-induced precipitation of alpha 2M-CH3NH2 or alpha 2M-trypsin is prevented by acylation of the protein employing the histidine-specific reagent diethylpyrocarbonate (DEP). The Zn2+-induced precipitation of alpha 2M-trypsin is prevented by acylation of the preformed alpha 2M-trypsin complex or by the reaction of acylated native alpha 2M with trypsin. Acylation of alpha 2M by treatment with DEP results in the modification of 13.5 histidyl residues per subunit of either native alpha 2M or alpha 2M-CH3NH2. Subsequent treatment with hydroxylamine reverses the modification of 10.5 histidyl residues per subunit in each protein preparation. These results indicate that histidyl residues are involved in the Zn2+-induced precipitation of alpha 2M-proteinase or alpha 2M-CH3NH2 complexes, and that these residues are accessible to extensive protein-metal interactions only after alpha 2M has undergone a major conformational change. These appear to be the same histidyl residues which, upon acylation by DEP, are responsible for recognition of alpha 2M-proteinase complexes by the acyl-low-density-lipoprotein cell surface receptor (S. V. Pizzo, P. A. Roche, S. R. Feldman, and S. L. Gonias (1986) Biochem. J. 238, 217-225).
D000215:Acylation; D011232:Chemical Precipitation; D004047:Diethyl Pyrocarbonate; D010450:Endopeptidases / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D006657:Histones; D011485:Protein Binding; D013057:Spectrum Analysis; D015032:Zinc; D000511:alpha-Macroglobulins / Q000378:metabolism*
D006657:Histones; D000511:alpha-Macroglobulins; D006639:Histidine; D010450:Endopeptidases; D015032:Zinc; D004047:Diethyl Pyrocarbonate