Western Blot Protein C functions as an anticoagulant by inhibiting factors V and

Question

Western Blot Protein C functions as an anticoagulant by inhibiting factors V and VIIl. People who have a mutation in this gene typically suffer from inappropriate blood clot formation in veins. There are 2 main types of mutations that lead to Protein C deficiency. Type I mutations lead to low levels of Protein C while type Il mutations do not effect Protein C levels, but instead affect its function. We had several patients that we knew had Protein C deficiency. We useda Western Blot technique to determine whether they were suffering from a type l or type Il deficiency and obtained the following results. a. (2 pts) What tissue would we isolate to perform this test? b. (2 pts) Are we testing for DNA, RNA, or protein? c. (2 pts) Which type of deficiency does patient 1 have? d. (2 pts) Which type of deficiency does patient 2 have? e. (2 pts) Give me an idea of how we could directly test for a type Il deficiency. Con P1 P2

Explanation / Answer

a. Since protein C is involved in blood clotting, the sample must be blood.

b. The western blotting technique is used to identify and quantitate proteins.

c. Type II mutation. Patient 1 has a normal level of protein. But the produced protein is non-functional.

d. Type I mutation. Patient 2 has a mutation that reduces the protein level.

e. We can isolate the protein from the patient sample and check for in-vitro blood clotting assay.

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