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. 2002 Oct;83(4):2248–2258. doi: 10.1016/S0006-3495(02)73985-4

Binding and relaxometric properties of heme complexes with cyanogen bromide fragments of human serum albumin.

Enrico Monzani 1, Maria Curto 1, Monica Galliano 1, Lorenzo Minchiotti 1, Silvio Aime 1, Simona Baroni 1, Mauro Fasano 1, Angela Amoresano 1, Anna Maria Salzano 1, Piero Pucci 1, Luigi Casella 1
PMCID: PMC1302313  PMID: 12324442

Abstract

The spectroscopic and reactivity properties of hemin complexes formed with cyanogen bromide fragments B (residues 1-123), C (124-298), A (299-585), and D (1-298) of human serum albumin (HSA) have been investigated. The complex hemin-D exhibits binding, spectral, circular dichroism, and reactivity characteristics very similar to those of hemin-HSA, indicating that fragment D contains the entire HSA domain involved in heme binding. The characteristics of the other hemin complexes are different, and a detailed investigation of the properties of hemin-C has been carried out because this fragment contains the HSA binding region of several important drugs. Hemin-C contains a low-spin Fe(III) center, with two imidazole ligands, but the complex undergoes a reversible structural transition at basic pH leading to a high-spin, five-coordinated Fe(III) species. This change determines a marked increase in the relaxation rate of water protons. Limited proteolysis experiments and mass spectral analysis carried out on fragment C and hemin-C show that the region encompassing residues Glu-208 to Trp-214 is protected from activity of proteases in the complex and, therefore, is involved in the interaction with hemin. A structural model of fragment C enables us to propose that His-242 and His-288 are the axial ligands for the Fe(III) center.

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