Mycobacterium tuberculosis에서 유래한 ADK의 발현, 정제 그리고 결정화 = Expression, Purification and Crystallization of Adenylate Kinase from Mycobacterium tuberculosis

목차 (Table of Contents)

• 1. Introduction 1
• 1.1 Tuberculosis 1
• 1.2 Nucleotide and NMPK (Nucleoside monophosphate kinase) superfamily 3
• 1.3 Adenylate kinase (ADK) 8
• 1.4. Crystal structure of Adenylate kinase (ADK) bound to nucleotides and nucleotide-free state 9
• 2. Materials and Methods 11
• 2.1. Protein expression 11
• 2.2. Crude extract preparation 13
• 2.3. Purification of ADK protein with a high quality 15
• 2.4. Enzyme kinetics 17
• 2.5. Crystallization 19
• 2.6. HPLC assay 21
• 2.7. Data collection and processing of ADK 22
• 3. Results 23
• 3.1. Overexpression and pre-purification of ADK 23
• 3.2. Purification of ADK 25
• 3.3. Kinetics of ADK 27
• 3.4. Crystallization of ADK 29
• 3.5. HPLC assay of ADK crystal 31
• 3.6. Preliminary X-ray data of ADK 33
• 4. Discussion 36
• 5. References 38