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Fig. 6 | Molecular Medicine

Fig. 6

From: Platelet Glycoprotein Ib: A Zinc-Dependent Binding Protein for the Heavy Chain of High-Molecular-Weight Kininogen

Fig. 6

Photoaffinity labeling of HK binding proteins on the surface of platelets. Twenty nanometers of biotin-modified HK was interacted with platelets in the presence or absence of 50 µM zinc. A separate control includes all reagents in the absence of HK. After photoactivation, cell lysis, and reduction with dithiothreitol, the proteins were examined by 10% SDS gel electrophoresis with development of bands with alkaline phosphatase conjugated avidin. (A) Lane 1, labeled protein in the presence of HK plus zinc; lane 2, labeled protein in the presence of HK without zinc; lane 3, a control in the absence of HK. In each instance, the proteins were isolated by avidin-sepharose affinity chromatography and examined by 7% SDS gel electrophoresis. (B) Lane 1, HK-interacting proteins in the presence of zinc; lane 2, Western blot with anti-GPIb of material in lane 1; lane 3, Western blot with anti-GP1b of interacting proteins in the absence of zinc.

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