結構生物學期中報告

1. 結構生物學期中報告 Speaker: 葉雅如 M93360018 04/00/26

2. Paper Calcium-pump inhibitors induce functional surface expression of ∆F508-CFTR protein in cystic fibrosis epithelial cells MARIE E. EGAN1,3, JUDITH GLÖCKNER-PAGEL3, CATHERINE A. AMBROSE1, PAULA A. CAHILL1,LAMIKO PAPPOE1, NAOMI BALAMUTH1, EDWARD CHO3, SUSAN CANNY3, CARSTEN A. WAGNER3,JOHN GEIBEL2,3 & MICHAEL J. CAPLAN3 Departments of 1Pediatrics, 2Surgery and 3Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut, USA J.G.-P. and C.A.A. contributed equally to this study. Correspondence should be addressed to M.J.C.; email: michael.caplan@yale.edu NATURE MEDICINE • VOLUME 8 • NUMBER 5 • MAY 2002 .485-492

3. PDB ID • Title : Cystic Fibrosis Transmembrane Conductance Regulator: Solution Structures Of Peptides Based On The Phe508 Region, The Most Common Site Of Disease-Causing -F508 Mutation • Classification :Metal Transport • 1CKY • Compound Mol_Id: 1; Molecule: Cystic Fibrosis Transmembrane Conductance Regulator (Cftr); Chain: A; Fragment: P26, F508 Mutation Region; Synonym: P26 In H2O; Engineered: Yes • 1CKZ • Compound Mol_Id: 1; Molecule: Cystic Fibrosis Transmembrane Conductance Regulator (Cftr); Chain: A; Fragment: P25, F508 Mutation Region; Synonym: P25 In H2O; Engineered: Yes

4. Introduction • Most cases of cystic fibrosis (CF), a common inherited disease of epithelial cell origin, are caused by the deletion of Phe508 located in the first nucleotide-binding domain (NBF1) of the protein called CFTR (cystic fibrosis transmembrane conductance regulator). • The most common mutation in cystic fibrosis, ∆F508, results in a cystic fibrosis transmembrane conductance regulator (CFTR) protein that is retained in the endoplasmic reticulum (ER). • Retention is dependent upon chaperone proteins, many of which require Ca++ for optimal activity. Interfering with chaperone activity by depleting ER Ca++ stores might allow functional ∆F508-CFTR to reach the cell surface.

5. Introduction A chloride channel • Figure 1 Model of CFTR. The model shows the two transmembrane-spanning domains (TMS1 and TMS2), the two nucleotide binding domains (NBF1 and NBF2), and the regulatory domain (R). The location of NBF1 peptides P25 and P26 used in this study is also indicated. Biochemistry,38 (23), 7453 -7461, 1999. 10.1021/bi9903603 S0006-2960(99)00360-8

6. Paper’s main question • 利用chaperone activity by depleting ER Ca++ stores,論文中以抑制劑來抑制the ER Ca++ pump 用以降低chaperone activity活性 使有功能性的 Δphe 508從ER釋放至細胞膜上 • 觀察代表性序列p26/p25的結構差異性 來分析為什麼會被chaperone辨識，並以此來考量治療性化學藥劑的使用。

7. Aim • 觀察p25/p26序列結構的呈現 • 針對Δphe 508缺失位置所影響的改變作探討

8. Result P26: 1cky / P25: 1ckz • Shap change

9. Result P26: 1cky / P25: 1ckz • Ramachandran Plot

10. Result • molecular surface P26: 1cky

11. Result • molecular surface P25: 1ckz

12. Result • Trimers P26: 1cky/P25: 1ckz

13. 接下來的目標 • 分析其他蛋白質與p25/26是否有保留性序列存在,是否有演化上的關係存在 • 其他蛋白質中包含p25/26相似的序列時,是否會造成該蛋白質立體結構改變

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