Individual work. student of _____ group. Lesson 3. Methods of enzymes investigation.

Individual work

student of _____ group

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^(name)

Lesson 3. Methods of enzymes investigation.

1. Name the methods of purification and separation of proteins, specific feathers of which are described below.

А. particles with different molecular weight move with different speed in column, which contains gel bids with different diameter’s of pores.

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B. substances with high molecular weight cannot penetrate the semipermeable membrane.

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C. substances are separated by degree of affinity for stationary or mobile faze.

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D. substances are separated by electric field in gradient of pH in dependence from pI level.

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2. What level of protein molecule organisation can be investigated by this method? Why the ending amino acids have to be marked?

ending amino acid

marking

polypeptide chain

marker

hydrolisis

3. Your task is to investigate primary structure of blood serum unknown protein. This protein moves with β-globulins during electrophoresis. Suggest your scheme of separation, purification, and determination of amino acid’s sequence of this molecule (write scheme of experiment: methods and their sequence).

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4.1. What physico-chemical properties of molecules are used for separation by electrophoresis?

4.2. Five fractions are found after paper electrophoresis of blood serum proteins, but near twenty fractions are found after polyacrylamid gel electrophoresis. Why?

5. What method of protein separation is based on high affinity of separating molecules to specific ligands, which are present in column (for example: separation of antigens on the column with antibodies)?

А) electrophoresis

B) ion exchange chromatography

C) gel – filtration

D) affinity chromatography

Е) salting out

6. What modification of iso-electric focusing method was proposed by Professor Troitskiy G. V.?

7. Fill in the scheme:

8. What is spectropolyarimetria?

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9. Fill in the scheme:

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