Circular dichroism studies of myoglobin and leghemoglobin

• Published in: Biochemistry (Easton) Vol. 14; no. 23; pp. 5141 - 5149
• Main Authors: Nicola, N. A, Minasian, E, Appleby, C. A, Leach, S. J
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 18.11.1975
• Subjects: Animals; Apoproteins; Binding Sites; Carbon Monoxide; Circular Dichroism; Cyanides; Glycine max; Heme - analysis; Hemeproteins; Leghemoglobin; Myoglobin; Nicotine; Oxidation-Reduction; Oxygen; Protein Binding; Protein Conformation; Species Specificity; Spectrophotometry; Spectrophotometry, Ultraviolet; Whales
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00694a019

The circular dichroism spectra of leghemoglobin a from the root nodules of soybean have been compared with those for sperm whale myoglobin in the fat- and near-ultraviolet and the Soret and visible regions of the spectrum. Circular dichroism spectra in the far-ultraviolet show that the leghemoglobins all have a high alpha-helix content (soybean leghemoglobin a, 55%) regardless of the nature of bound ligands and oxidation or spin state of the heme iron. The known sequence homologies with mammalian hemoglobins may therefore be reflected in conformational homologies as suggested by the x-ray studies of Vainshtein et al. ((1975) Nature (London) 254, 163-164) on lupin leghemoglobin. Removal of the heme moiety decreases helicity by only 9% for leghemoglobins, compared with 23% for myoglobin. This, the much smaller heme contribution to the near-ultraviolet circular dichroism than in myoglobin, and the greater accessibility of the heme moiety to aqueous solvent (Nicola et al. (1974), Proc. Aust. Biochem. Soc. 7, 21) suggest that the association between heme and protein is much weaker in leghemoglobins than in myoglobin. The aromatic Soret and visible circular dichroism spectra for all derivatives of leghemoglobin are opposite in sense to those for myoglobin, showing that the patterns of protein side chain contacts with the heme are different in the two classes of heme proteins. There is strong evidence that one of the two tryptophans whose identity and structural role in myoglobin is known, is present also in plant leghemoglobins, hydrogen-bonded and in a similar nonpolar environment whether heme is present or not. The above findings help to explain the remarkably high oxygen affinity and some other ligand-binding properties of leghemoglobins which differ from those of myoglobin.