Electron microscopy studies of alpha 2-macroglobulin conformational intermediates obtained by derivatization with cis-dichlorodiammineplatinum (II).
The structure of human alpha 2-macroglobulin (alpha 2M) after reaction with cis-dichlorodiammineplatinum (II) (cis-DDP) was studied by electron microscopy. The cis-DDP stabilized a novel conformation of the native inhibitor resembling a doughnut surrounded by two, three, of four well defined spherules. When only two spherules were present, these structures were usually oriented on opposite sides of the doughnut. The protein region joining a spherule to the central structure did not include sufficient mass to exclude stain and was, therefore, invisible. Other images showed spherules that were partially superimposed on the doughnut. A comparison of many molecules suggested great flexibility of the peripheral spherules relative to the central structure. The cis-DDP prevented complete conformational change when the alpha 2M was reacted with trypsin. The products of this reaction included apparent conformational intermediates. These intermediates most closely resembled either native alpha 2M or the well established "H" structure of alpha 2M-proteinase, depending on the initial conditions used to modify the alpha 2 M with cis-DDP. When cis-DDP-treated alpha 2M was reacted with trypsin, purified by chromatography and subsequently treated with diethyldithiocarbamate, complete conformational change was observed. Based on an analysis of the alpha 2M structural intermediates obtained using the chemical modification procedures described here, a new model of alpha 2M conformational change was developed. We postulate that conformational change initially involves contraction of the peripheral spherules towards the central doughnut. These spherules then unfold and elongate in the perpendicular direction to form the lateral walls of the proteinase transformed alpha 2M H structure.
Gonias
S L
SL
Department of Pathology, University of Virginia Health Sciences Center, Charlottesville 22908.
Figler
N L
NL
D002945:Cisplatin; D003432:Cross-Linking Reagents; D004050:Ditiocarb; D006801:Humans; D006868:Hydrolysis; D008854:Microscopy, Electron; D015394:Molecular Structure; D011487:Protein Conformation / Q000187:drug effects*; D014357:Trypsin; D000511:alpha-Macroglobulins / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*
D003432:Cross-Linking Reagents; D000511:alpha-Macroglobulins; D004050:Ditiocarb; D014357:Trypsin; D002945:Cisplatin