Abstract

This research will develop capillary electrophoresis for studying fundamental interactions in protein biochemistry. Its focus is on """"""""protein charge ladders""""""""--families of proteins prepared by acetylation of lysine amino groups, and that differ in their net charge; these families (the """"""""rungs"""""""") of the ladders separate in capillary electrophoresis into a set or regularly spaced peaks on the basis of net charge. Study of the interactions of protein charge ladders with charged ligands, with buffers, and with surfactants gives detailed information about these interactions. Charge ladders were originally developed for studying electrostatic interactions; this work will extend the study to the investigation of hydrophobic interactions. The information developed in this research will help in understanding and manipulating a wide range of fundamental biochemical interactions. The research will also use the techniques of soft lithography to develop new types of microfluidic systems for use in capillary electrophoresis and other fluidic applications. The research will provide understanding useful in rationalizing and predicting interactions between proteins and ligands, and ultimately in the rational design of drugs.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM051559-10
Application #
6770026
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Edmonds, Charles G
Project Start
1997-12-01
Project End
2006-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
10
Fiscal Year
2004
Total Cost
$244,500
Indirect Cost

  Institution

Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138

  Publications

Shaw, Bryan F; Schneider, Gregory F; Whitesides, George M (2012) Effect of surfactant hydrophobicity on the pathway for unfolding of ubiquitin. J Am Chem Soc 134:18739-45
Shaw, Bryan F; Schneider, Gregory F; Arthanari, Haribabu et al. (2011) Complexes of native ubiquitin and dodecyl sulfate illustrate the nature of hydrophobic and electrostatic interactions in the binding of proteins and surfactants. J Am Chem Soc 133:17681-95
Lee, Andrew; Mirica, Katherine A; Whitesides, George M (2011) Influence of fluorocarbon and hydrocarbon acyl groups at the surface of bovine carbonic anhydrase II on the kinetics of denaturation by sodium dodecyl sulfate. J Phys Chem B 115:1199-210
Lee, Andrew; Tang, Sindy K Y; Mace, Charles R et al. (2011) Denaturation of proteins by SDS and tetraalkylammonium dodecyl sulfates. Langmuir 27:11560-74
Mecinovic, Jasmin; Snyder, Phillip W; Mirica, Katherine A et al. (2011) Fluoroalkyl and alkyl chains have similar hydrophobicities in binding to the ""hydrophobic wall"" of carbonic anhydrase. J Am Chem Soc 133:14017-26
Snyder, Phillip W; Mecinovic, Jasmin; Moustakas, Demetri T et al. (2011) Mechanism of the hydrophobic effect in the biomolecular recognition of arylsulfonamides by carbonic anhydrase. Proc Natl Acad Sci U S A 108:17889-94
Lee, Andrew; Whitesides, George M (2010) Analysis of inorganic polyphosphates by capillary gel electrophoresis. Anal Chem 82:6838-46
Shaw, Bryan F; Arthanari, Haribabu; Narovlyansky, Max et al. (2010) Neutralizing positive charges at the surface of a protein lowers its rate of amide hydrogen exchange without altering its structure or increasing its thermostability. J Am Chem Soc 132:17411-25
Krishnamurthy, Vijay M; Kaufman, George K; Urbach, Adam R et al. (2008) Carbonic anhydrase as a model for biophysical and physical-organic studies of proteins and protein-ligand binding. Chem Rev 108:946-1051
Schneider, Gregory F; Shaw, Bryan F; Lee, Andrew et al. (2008) Pathway for unfolding of ubiquitin in sodium dodecyl sulfate, studied by capillary electrophoresis. J Am Chem Soc 130:17384-93

Showing the most recent 10 out of 38 publications