SAXS Reveals Solution Structures of Gelsolin G2-G3 Fragment and Actin-Peptide Complex

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研究生：	黃頌喬 Huang, Song-Chiao
論文名稱：	SAXS Reveals Solution Structures of Gelsolin G2-G3 Fragment and Actin-Peptide Complex
指導教授：	張石麟 Chang, Shih-Lin 黃玉山 Huang, Yu-Shan
口試委員:	蘇雲良湯茂竹
學位類別：	碩士 Master
系所名稱：	理學院 - 先進光源科技學位學程 Degree Program of Science and Technology of Synchrotron Light Source
論文出版年：	2012
畢業學年度：	100
語文別：	英文
論文頁數：	53
中文關鍵詞：	小角度X光散射、蛋白質水溶液結構
外文關鍵詞：	SAXS, Gelsolin, Solution Structures, Actin-Peptide complex
相關次數：	點閱：3 下載：0
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ABSTRACT

Gelsolin is a key regulator for action dynamics which has six homologous domains notated from G1 to G6. It has the ability for severing, disassembling, or capping the actin filaments. Reviewing relative literatures, there are two different reported structures of G2-G3 fragment for gelsolin. In this report, small-angle X-ray scattering (SAXS) has been applied to analyze structures of G2-G3 fragments from two different constructs. Each construct is composed of two tertiary domains G2 and G3, a flexible header and linker. The difference between the two constructs is the length of the flexible header. Both solution structures of the two constructs have been revealed by using the ab-initio method and rigid-body refinement. A comparison of the crystal and solution structures suggests that they are slightly different from each others. Moreover, the flexible header might interfere relative orientations of G2 and G3 domains.
The other subject of interest is to reveal the solution structure of the actin-peptide complex, which is composed of three actins and one peptide. Actin participates in many important cellular processes, including muscle contraction, cell motility, maintenance of cell, and cell signaling. Peptide is a polymer of several amino acids. The two possible solution structures of actin-peptide complex have been reported by using SAXS analysis.

Key words: SAXS, gelsolin, actin filament, protein solution structure

Content
Chapter 1 Introduction    1
Chapter 2 SAXS Theory and Principle    3
1 Principle of Small-Angle X-Ray Scattering    3
3 Concept of Ab-Initio Method for Shape Building in Low-Resolution    8
3.1 Ab-Initio Method by Densely Packed Bead Model    9
3.2 Ab-Initio Method by Dummy Residue Model    10
4 Concept of Rigid-Body Refinement for Detailed Model Building    12
Chapter 3 Experiment and Instrumentation    13
1 Introduction to Gelsolin    14
2 SAXS Analysis for Gelsolin Fragment (Construct 1)    15
2.1 Introduction to Solution Construct 1    15
2.2 SAXS Profile by Guinier Plot and Data Reduction    17
2.3 Pair Distance Distribution Function    18
2.4 Ab-Initio Method with Bead Model    19
2.5 Ab-Initio Method with Dummy Residue Model    21
2.6 Rigid-Body Refinement for High-Resolution Structure    23
2.7 Structures Alignment for Model Reliability    25
2.8 The Solution Structure Align with Two Crystallographic Structures    26
3 SAXS Analysis for Gelsolin Fragment (Construct 2)    27
3.1 Introduction to Gelsolin Fragment (Construct 2)    27
3.2 SAXS Profile by Guinier Plot and Data Reduction    28
3.4 Ab-Initio Method with Bead Model    30
3.5 Ab-Initio Method with Dummy Residue Model    32
3.6 Rigid-Body Refinement for High-Resolution Structure    34
3.7 Structures Alignment for Model Reliability    36
3.8 The Solution Structure Align with Two Crystallographic Structures    37
4 Discussion    38
Chapter 5 Result and Discussion for Actin-Peptide    39
1 Introduction to Actin-Peptide    39
2 SAXS Analysis for Actin-Peptide    41
2.1 SAXS Profile by Guinier Plot and Data Reduction    41
2.2 Pair Distance Distribution Function    42
2.3 Ab-Initio Method with Bead Model    43
2.4 Ab-Initio Method with Dummy Residue Model    45
2.5 Rigid-Body Refinement for High-Resolution Structure    47
2.6 Structure Alignment for Model Reliability    49
3 Discussion and Future Work    51
References    52

                                

References
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