Abstract

Polytopic helical integral membrane proteins are an important class of proteins that have been understudied because of technical issues regarding their expression, purification, and crystallization. Total chemical protein synthesis represents a potential route to these important proteins, but is currently hampered by insolubility of the transmembrane peptides required for protein assembly. To address this issue, we propose to develop a general method to render transmembrane peptides soluble for the manipulations necessary in chemical protein synthesis. We will then synthesize a 7 TM integral membrane protein to demonstrate the viability of chemical membrane protein synthesis. Establishing routine synthetic access to integral membrane proteins will provide tools to study their mechanisms in detail not currently available to molecular biology, which will eventually lead to a better understanding of how these proteins function in normal and pathological states.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM075993-03
Application #
7265325
Study Section
Special Emphasis Panel (ZGM1-PPBC-3 (MP))
Program Officer
Chin, Jean
Project Start
2005-09-23
Project End
2010-07-31
Budget Start
2007-08-01
Budget End
2008-07-31
Support Year
3
Fiscal Year
2007
Total Cost
$267,821
Indirect Cost

  Institution

Name
University of Chicago
Department
Biochemistry
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637

  Publications

Liu, Suhuai; Pentelute, Brad L; Kent, Stephen B H (2012) Convergent chemical synthesis of [lysine(24,38,83)] human erythropoietin. Angew Chem Int Ed Engl 51:993-9
Yeates, Todd O; Kent, Stephen B H (2012) Racemic protein crystallography. Annu Rev Biophys 41:41-61
Mandal, Kalyaneswar; Kent, Stephen B H (2011) Total chemical synthesis of biologically active vascular endothelial growth factor. Angew Chem Int Ed Engl 50:8029-33
Sohma, Youhei; Hua, Qing-Xin; Whittaker, Jonathan et al. (2010) Design and folding of [GluA4(ObetaThrB30)]insulin (""ester insulin""): a minimal proinsulin surrogate that can be chemically converted into human insulin. Angew Chem Int Ed Engl 49:5489-93
Banigan, James R; Mandal, Kalyaneswar; Sawaya, Michael R et al. (2010) Determination of the X-ray structure of the snake venom protein omwaprin by total chemical synthesis and racemic protein crystallography. Protein Sci 19:1840-9
Pentelute, Brad L; Mandal, Kalyaneswar; Gates, Zachary P et al. (2010) Total chemical synthesis and X-ray structure of kaliotoxin by racemic protein crystallography. Chem Commun (Camb) 46:8174-6
Mandal, Kalyaneswar; Pentelute, Brad L; Tereshko, Valentina et al. (2009) Racemic crystallography of synthetic protein enantiomers used to determine the X-ray structure of plectasin by direct methods. Protein Sci 18:1146-54
Mandal, Kalyaneswar; Pentelute, Brad L; Tereshko, Valentina et al. (2009) X-ray structure of native scorpion toxin BmBKTx1 by racemic protein crystallography using direct methods. J Am Chem Soc 131:1362-3
Sohma, Youhei; Kent, Stephen B H (2009) Biomimetic synthesis of lispro insulin via a chemically synthesized ""mini-proinsulin"" prepared by oxime-forming ligation. J Am Chem Soc 131:16313-8
Pentelute, Brad L; Gates, Zachary P; Dashnau, Jennifer L et al. (2008) Mirror image forms of snow flea antifreeze protein prepared by total chemical synthesis have identical antifreeze activities. J Am Chem Soc 130:9702-7

Showing the most recent 10 out of 15 publications