title: Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins.
abstract: Spectroscopic measurement of protein concentration requires knowledge of the value of the relevant extinction coefficient. If the amino acid composition of a protein is known, however, extinction coefficients can be calculated approximately, provided that the values of the molar absorptivities for tryptophan and tyrosine residues in the protein are known. We have applied a matrix linear regression procedure and a mapping of average absolute deviations between experimental and calculated values to find molar extinction coefficients (epsilon M, 1 cm, 280 nm) of 5540 M-1 cm-1 for tryptophan and 1480 M-1 cm-1 for tyrosine residues in an "average" protein, as defined by a set of experimentally determined extinction coefficients for more than 30 proteins. Use of these values provides a significant improvement in extinction coefficient estimation over that obtained with the commonly used values obtained from solutions of model compounds in guanidine-HCl. The consistency of these results when compared to the large deviations often observed between experimentally determined extinction coefficients suggest that this method may offer acceptable accuracy in the initial estimation of molar absorptivities of globular proteins.
authors.affiliation: Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486.
authors.forename: H
authors.initials: H
authors.lastname: Mach
authors.forename: C R
authors.initials: CR
authors.lastname: Middaugh
authors.forename: R V
authors.initials: RV
authors.lastname: Lewis
mesh_terms: D000818:Animals; D004798:Enzymes / Q000737:chemistry*; D006801:Humans; D011506:Proteins / Q000737:chemistry*; D013057:Spectrum Analysis; D014364:Tryptophan / Q000737:chemistry*; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D004798:Enzymes; D011506:Proteins; D014443:Tyrosine; D014364:Tryptophan