AIM MATERIALS AND METHODS ABSTRACT INTRODUCTION INDEX

INDEX

ABSTRACT

INTRODUCTION

1.

Glutathione

1.3 Biological functions of glutathion………...9

2.

γ

-Glutamyltranspherase

2.1. γGT localization…………..………...14

2.2. γGT Structure ………..………15

2.3. γGT binding site and catalytic site………...…15

2.4. γGT Structure biosynthesis and gene expression……….…17

2.5. Physiological function of γGT……….…18

2.6. γGT pro-oxidant action………19

2.7. γGT in human tumor………20

2.8. γGT and drug resistance………...…21

2.9. Association of serum γGT with several human diseases…….……22

3.

γ

-Glutamyltranspherase related enzyme

3.1Localization and expression of γGT-rel enzyme………...…....23

3.2 γGT-rel structure………..…………..…………..…… 24

3.3 Mechanism of action and main physiological substrates…………..25

3.4 γGT-rel pro-oxidant action………..…………..……...27

3.5 Roles of γGT rel in the respiratory system………..….……27

AIM

MATERIALS AND METHODS

2. Methods

2.1 Assay with chromogenic substrate L-γ-glutamyl-p-nitroanilide..…31

2.2 Assay with GSH as substrate………..…………..………31

2.3 Determination of protein concentration………..……..…32

2.4 Preparation of crude extracts………..…………..….33

2.5 Ultracentrifugation……….…..…………..…………..….34

2.6 Ammonium sulphate precipitation………..…………..34

2.7 Hydrophobic interaction chromatography………34

2.8 Gel filtration………..…………..…………..…………35

2.9 Affinity chromatography………..…………..………...………35

RESULTS

2. Study on the identification and purification of γGT-rel from bovine spleen………..…………..…………..…………..………..45

3. Development of a colorimetric method for the determination of glutathione…………..………..…………..…53

DISCUSSION AND CONCLUSIONS