Question

which of the following amino acids are used to measure the protein concentration by UV? A. His B. Pro C. Trp D. Phe E. Tyr

Answer 1

Answer:

C. Trp D. Phe E. Tyr

Explanation:

The concentration of a protein has a direct relation with absorbance of the protein in a UV spectrophotometer. The formula which relates concentration with absorbance is described as under:

                A = ∈ x c x l

where, A = Absorbance

            ∈ = Molar extinction co-efficient

            c  = Concentration of absorbing species i.e. protein

             l  = Path length of light

       

Tryptophan (Trp), phenylalanine (Phe ) and tyrosine (Tyr) are three aromatic amino acids which are used to measure protein concentration by UV. It is mainly because of tryptophan (Trp), protein absorbs at 280 nm which gives us an idea of protein concentration during UV spectroscopy.

The table depicting the wavelength at which these amino acids absorb and their respective molar extinction coefficient is as under:

Amino acid                Wavelength            Molar extinction co-efficient (∈)

Tryptophan                     282 nm                        5690

Tyrosine                          274 nm                        1280

Phenylalanine                 257 nm                        570

In view of table above, we can easily see that Molar extinction co-efficient (∈) of Tryptophan is highest amongst all these 3 amino acids that is why it dominates while measuring concentration.