Resolution of the individual tryptophanyl contributions to the near-ultraviolet dichroic activity of apomyoglobin.
The individual tryptophanyl contributions to the near-ultraviolet dichroic activity of apomyoglobin in its native conformation have been resolved. This was accomplished by comparing the spectra of two classes of apomyoglobin with different aromatic residue contents and observing the effect of a specific modification of indole residues. The circular dichroism (CD) spectra of apomyoglobins containing two tryptophanyl residues, i.e. Trp A-5 and A-12, show the presence of a positive peak centered at 292 nm, attributable to indolic chromophore, which is missing in the CD spectrum of tuna apomyoglobin possessing only Trp A-12. Moreover, the specific modification of Trp A-5 by 2-hydroxy-5-nitrobenzyl bromide is shown by the lack of the 292 nm peak and the appearance of a positive band at longer wavelength. The pH dependence of the position of this band suggests that it arises from the 2-hydroxy-5-nitrobenzyl moiety. The results suggest that Trp A-12 does not substantially contribute to the optical activity in the near ultraviolet.
Sirangelo
I
I
Dipartimento di Biochimica e Biofisica, Seconda Università di Napoli, Italy.
Irace
G
G
Bismuto
E
E
D000818:Animals; D001059:Apoproteins / Q000737:chemistry*; D002942:Circular Dichroism; D009211:Myoglobin / Q000737:chemistry*; D010777:Photochemistry; D013056:Spectrophotometry, Ultraviolet; D014364:Tryptophan / Q000737:chemistry
D001059:Apoproteins; D009211:Myoglobin; C010857:apomyoglobin; D014364:Tryptophan