Open quaternary structure of the hagfish proteinase inhibitor with similar properties to human alpha-2-macroglobulin.
A homologous protein to human plasma alpha-2-macroglobulin (alpha-2-M) was purified from the blood plasma of hagfish (Eptatretus buergeri) and its structure and function were studied. The hagfish protein inhibited several proteinases and its inhibitory activity was blocked with methylamine as in the case of human alpha-2-M. The molecular weight and sedimentation coefficient of the hagfish inhibitor were 390,000 +/- 20,000 and 11.0 S, respectively, as determined by sedimentation studies. The frictional ratio calculated from these parameters was 1.75. The Stokes radius estimated from HPLC gel chromatography was 8.8-8.9 nm, which was similar to that of human alpha-2-M despite the fact that the hagfish inhibitor was only one-half as large as human alpha-2-M in molecular weight. The hagfish inhibitor was expected to be more asymmetric and/or more hydrated than the human inhibitor. The electron micrographs of the negatively stained hagfish inhibitor showed that it had an open, rectangular quaternary structure of 15 +/- 1.5 X 19 +/- 2 nm in which two semiglobular units were located at the two shorter sides with a gap of 8 +/- 1 nm in width. Each semiglobular unit had an approximate width of 5 +/- 0.5 nm. The thickness of the unit was estimated to be 3 to 3.5 nm from the result of fixed-angle shadowing experiments. Although the two semiglobular units must be connected by some structure, very little material could be seen between them. Such an open quaternary structure may explain the high frictional ratio and large Stokes radius of this protein. The structural change of the inhibitor after reaction with proteinases or methylamine could be detected by electron microscopy and gel chromatography.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D005399:Fishes / Q000097:blood*; D006196:Hagfishes / Q000097:blood*; D006801:Humans; D008968:Molecular Conformation; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000097:blood
D000596:Amino Acids; D011480:Protease Inhibitors; D000511:alpha-Macroglobulins