Using Protein Charge Ladders to Estimate the Effective Charges and Molecular Weights of Proteins in Solution

Analytical Chemistry

Volumn 69, Issue 4, 1997, Pages 575-580

  Gao, Jinming ^a   Whitesides, George M ^a  

^a HARVARD UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ACETIC ANHYDRIDE; PROTEIN;

ARTICLE; CHEMISTRY; MOLECULAR WEIGHT; POLYACRYLAMIDE GEL ELECTROPHORESIS; PROTEIN CONFORMATION; SOLUTION AND SOLUBILITY;

ACETIC ANHYDRIDES; ELECTROPHORESIS, POLYACRYLAMIDE GEL; MOLECULAR WEIGHT; PROTEIN CONFORMATION; PROTEINS; SOLUTIONS;

EID: 0031568682     PISSN: 00032700     EISSN: None     Source Type: Journal    
DOI: 10.1021/ac9608073     Document Type: Article

References (27)

1. Grossman, P. D. Capillary Electrophoresis: Theory and Practice; Academic Press San Diego, CA, 1992; pp 111-132.
2. Compton, B. J.; O'Grady, E. A. Anal. Chem. 1991, 63, 2597-2602.
3. Engelhardt, H.; Beck, W.; Kohr, J.; Schmitt, T. Angew. Chem., Int. Ed. Engl. 1993, 32, 629-766.
4. Kuhr, W. G.; Monnig, C. A. Anal. Chem. 1992, 64, 389-407.
5. Novotny, M. V.; Cobb, K. A.; Liu, J. Electrophoresis 1990, 11, 735-749.
6. Karger, B. L.; Cohen, A. S.; Guttman, A. J. Chromatogr. 1989, 492, 585-614.
7. Gordon, M. J.; Huang, X.; Pentoney, S. L.; Zare, R. N. Science 1988, 242, 224-228.
8. Basak, S. K.; Ladisch, M. R. Anal. Biochem. 1995, 226, 51-58.
9. Rickard, E. C.; Strohl, M. M.; Nielsen, R. G. Anal. Biochem. 1991, 197, 197-207.
10. Abramson, H. A.; Moyer, L. S.; Gorin, M. H. Electrophoresis of Proteins and the Chemistry of Cell Surfaces; Reinhold Publishing Corp.: New York, 1942; pp 105-172.
11. The typical range of ionic strength that is compatible with CE analysis is between 1 mM and 1 M.
12. 0 is the permitivity of free space, k is the Boltzmann constant, and T is the absolute temperature. See ref 1.
13. Gao, J.; Mammen, M.; Whitesides, G. M. Science 1996, 272, 535-537.
14. Gao, J.; Mrksich, M.; Goraez, F. A.; Whitesides, G. M. Anal. Chem. 1995, 34, 3093-3100.
15. Gao, J.; Gomez, F.; Haerter, R.; Whitesides, G. M. Proc. Natl. Acad. Sci. U.SA. 1994, 91, 12027-12030.
16. Modification of Arg residues on a protein by phenylglyoxal will also yield a charge ladder with integral unit of charge difference: Takahashi. K. J. Biol. Chem. 1968, 243, 6171-6179. Roberts, C.; Córdon, E.; Whitesides, G. M., unpublished results.
17. Modification of Arg residues on a protein by phenylglyoxal will also yield a charge ladder with integral unit of charge difference: Takahashi. K. J. Biol. Chem. 1968, 243, 6171-6179. Roberts, C.; Córdon, E.; Whitesides, G. M., unpublished results.
18. Córdova, E.; Gao, J.; Whitesides, G. M. Anal. Chem., submitted.
19. Worthington, V. Worthington Enzyme Manual; Worthington Biochemical Corp,: Freehold, NJ, 1993.
20. Righetti, P. G.; Caravaggio, T. J. Chromatogr. 1976, 127, 1-28.
21. The peaks due to modification on the α-amino group of a protein are assigned by the difference of chemical reactivity of the α-amino group toward acetic anhydride compared to that of an ε-amino group. See ref 14.
22. The suggestion of differences in drag is a very much unproved hypothesis. If it is correct, it might be caused by the change of conformation of the protein upon modification.
23. α.
24. Creighton, T. E. PROTEINS Structures and Molecular Properties; W. H. Freeman and Co.: New York, 1993; p 266.
25. Nemoto, N.; Koike, A.; Osaki, K.; Koseki, T.; Doi, E. Biopolymers 1993, 33, 551-559.
26. Matsumoto, T.; Chiba, J. J. Chem. Soc., Faraday Trans. 1990, 86, 2877-2882.
27. P of ovalbumin as a function of concentration of dioxane in solution. The result demonstrates that added dixoane in the buffer did not change the state of aggregation of ovalbumin. These results suggest that the commercial sample of ovalbumin (obtained from Sigma) may already be irreversibly modified to exist primarily as a dimer in solution.