A simple and rapid method to study the association of the contact proteins of blood coagulation.
Native and reduced SDS polyacrylamide gel electrophoresis on the automated PhastSystem (Pharmacia) were used to demonstrate protein-protein binding interactions and structural changes during proteolytic activations of the proteins involved in contact activation. The "mobility shift" assay in native gels has been used to visualize the kinetics of activation of factor XII by dextran sulfate as well as the formation of kallikrein-cleaved high molecular weight kininogen. It shows the formation of prekallikrein-high molecular weight kininogen complexes and factor XII-dextran sulfate complex for the first time in gels. The use of automation makes this procedure fast and reproducible using nanogram amounts of protein in relatively short time.
Katcher
H L
HL
Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla 10595.
Samuel
M
M
Villanueva
G B
GB
D000595:Amino Acid Sequence; D016264:Dextran Sulfate / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005174:Factor XII / Q000378:metabolism*; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors; D007704:Kininogens / Q000378:metabolism*; D008969:Molecular Sequence Data; D011288:Prekallikrein / Q000378:metabolism*; D011485:Protein Binding; D012967:Sodium Dodecyl Sulfate
D007704:Kininogens; D012967:Sodium Dodecyl Sulfate; D005174:Factor XII; D016264:Dextran Sulfate; D011288:Prekallikrein; D007610:Kallikreins