Image processing of proteinase- and methylamine-transformed human alpha 2-macroglobulin. Localization of the proteinases.
Comparative x-ray scattering experiments and electron microscopic observations have been performed on native S-form, and on different F-forms of human plasma alpha 2-macroglobulin (alpha 2M), obtained by proteinase (chymotrypsin, plasmin, and thrombin) or methylamine treatment. Image processing of electron micrographs of the alpha 2M molecules transformed by chymotrypsin, plasmin, and methylamine displayed average images which could be compared. The proteinase-complex alpha 2M molecules exhibited the usual H-like structure, but the methylamine-inactivated ones showed a different organization, with almost no stain-excluding material in the central region of the molecule, which therefore presented a central cavity filled with stain. By subtracting average images of alpha 2M-methylamine from alpha 2M-chymotrypsin or alpha 2M-plasmin, a putative localization of the proteinases inside the alpha 2M molecule, very close to its center was revealed. The values of the radii of gyration for the S- and F-forms obtained by x-ray scattering were very different (78 and 67.7 A, respectively). All four scattering curves of the F-forms were comparable in shape and showed maxima and minima different from that of the S-form alpha 2M. Image processing of electron micrographs and x-ray scattering have provided independent results which indicate that a large cavity exists in the alpha 2M-methylamine molecule and that the proteinases might be located in a very central position inside the alpha 2M-proteinase molecules.
Boisset
N
N
Laboratoire de Biochimie Fondamentale, Tours, France.
Taveau
J C
JC
Pochon
F
F
Tardieu
A
A
Barray
M
M
Lamy
J N
JN
Delain
E
E
D002918:Chymotrypsin / Q000494:pharmacology*; D005341:Fibrinolysin / Q000494:pharmacology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008744:Methylamines / Q000494:pharmacology*; D008854:Microscopy, Electron; D011487:Protein Conformation; D013917:Thrombin / Q000494:pharmacology*; D014961:X-Ray Diffraction; D000511:alpha-Macroglobulins / Q000378:metabolism*
D008744:Methylamines; D000511:alpha-Macroglobulins; C027451:methylamine; D002918:Chymotrypsin; D013917:Thrombin; D005341:Fibrinolysin