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Calculation of protein extinction coefficients from amino acid sequence data
1Institute of Molecular Biology, University of Oregon, Eugene 97403.
Analytical Biochemistry
|November 1, 1989
Summary
Accurately determine protein concentration using UV spectroscopy by calculating molar extinction coefficients. This method uses amino acid composition, providing reliable results for proteins lacking experimental data.
Area of Science:
- Biochemistry
- Molecular Biology
- Spectroscopy
Background:
- Accurate protein concentration determination is crucial for studying molecular interactions.
- Experimental measurement of molar extinction coefficients is often infeasible for small protein quantities.
- UV spectroscopy is a common method for protein quantification, but requires accurate molar extinction coefficients.
Purpose of the Study:
- To develop a method for calculating accurate molar extinction coefficients for proteins at 280 nm.
- To enable reliable protein concentration determination from amino acid sequence data.
Main Methods:
- Calculating molar extinction coefficients based on amino acid composition.
- Utilizing known DNA sequences to derive amino acid composition and molecular weight.
- Calibrating the method against 18 well-characterized globular proteins.
Main Results:
- The developed method calculates accurate molar extinction coefficients (within +/- 5% for most proteins).
- This approach overcomes limitations of experimental measurements for proteins available in limited amounts.
Conclusions:
- The method provides a reliable way to estimate protein molar extinction coefficients from amino acid sequence.
- This facilitates accurate protein concentration measurements essential for biochemical and molecular biology studies.