Three-dimensional structures of the human alpha 2-macroglobulin-methylamine and chymotrypsin complexes.
The three-dimensional structures of chymotrypsin- and methylamine-treated negatively stained human alpha 2-macroglobulin have been determined by weighted back projection from electron microscope data. Projections of the reconstructions show good concordance with two-dimensional averages of both stained and frozen-hydrated molecules. The reconstructions reveal that the H-shaped front projection of the molecule is related to the smaller ellipsoidal end view by a rotation of 90 degrees about the crossbar (minor axis) of the H. This finding is in agreement with tilt studies. The reconstruction of the alpha 2-macroglobulin-methylamine reveals an hour-glass shaped void which is filled by the two proteinase molecules in the reconstruction of alpha 2-macroglobulin-chymotrypsin. Protein plugs which appear to block the exterior entrances to the cavity may function to prevent access of proteins to the encapsulated proteinase and serve to block its escape. Extensive thresholding of each reconstruction leaves a "backbone" consisting of two side-by-side rod-like structures, suggesting that this is the arrangement of the two protomeric units which form the molecule. Both structures show some departure from the expected symmetry. The asymmetries are robust features of the reconstructions and may reflect structurally asymmetric features of the transformation from the native to the chymotrypsin-treated form of the molecule.
Schroeter
J P
JP
Department of Pathology, University of Texas Health Science Center, Houston 77030.
Kolodziej
S J
SJ
Wagenknecht
T
T
Bretaudiere
J P
JP
Tapon-Bretaudiere
J
J
Strickland
D K
DK
Stoops
J K
JK
D001665:Binding Sites; D002918:Chymotrypsin / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008744:Methylamines / Q000737:chemistry; D008854:Microscopy, Electron; D008958:Models, Molecular; D015394:Molecular Structure; D011487:Protein Conformation; D000511:alpha-Macroglobulins / Q000737:chemistry / Q000648:ultrastructure*
D008744:Methylamines; D000511:alpha-Macroglobulins; C027451:methylamine; D002918:Chymotrypsin