Bovine hemoglobin as a potential source of hemoglobin-based oxygen carriers: crosslinking with bis (2,3-dibromosalycyl) fumarate
Fronticelli, C.; Sato, T.; Orth, C.; Bucci, E.
Biochimica et Biophysica Acta 874(1): 76-81
1986
ISSN/ISBN: 0006-3002 PMID: 3768378 Accession: 004854993
Reaction in anaerobic conditions of bovine hemoglobin with bis(2,3-dibromosalycyl)fumarate resulted in new derivatives with p50 in excess of 40 mmHg, as determined at 37.degree. C in 0.15 M Cl- at pH 7.4. Although the chromatographic preparations indicated some heterogeneity of the reacted material, the proteins obtained were homogeneous with regard to sedimentation velocity, which showed the presence of only nondissociable tetrameric species. SDS gel electrophoresis showed the presence of a new band with a mobility corresponding approximately to a molecular mass of 32 kDa, indicating the presence of covalent intramolecular crosslinks between subunit pairs. Chromatographic analyses indicated that both .alpha. and .beta. chains were chemically modified. The retention times in rats of the crosslinked hemoglobin was 10-times longer than that of untreated hemoglobin.