The Protein Journal

, Volume 26, Issue 4, pp 257-263

Temperature Dependent Soret Spectral Band Shifts Accompany Human CN-Mesohemoglobin Assembly

  • Priyani  V. FonsekaAffiliated withDepartment of Chemistry, University of MassachusettsColumbus Children’s Hospital Research Institute
  • , Gayathri VasudevanAffiliated withDepartment of Chemistry, University of MassachusettsDepartment of Chemistry, Wellesley College
  • , Lisa-Jo  Ann ClariziaAffiliated withDepartment of Chemistry, University of Massachusetts
  • , Melisenda  J. McDonaldAffiliated withDepartment of Chemistry, University of Massachusetts Email author 

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The interaction between human apohemoglobin A and CN-Mesohemin, a monomeric non-native heme derivative, was probed by Soret spectrophotometric titrations in 0.05 M potassium phosphate buffer, pH 7 at varied temperatures. Hypsochromic shifts in the absorbance maxima were observed at all temperatures below 10°C. First derivative spectroscopy of CN-Mesohemin titrations was used to provide further evidence of a spectral shift upon CN-Mesohemoglobin assembly. Findings of Soret Spectral shifts demonstrate a preference for the α chain heme site by CN-Mesohemin indicative of semi-α-hemoglobin intermediate formation. CN-Mesohemin, a derivative with peripheral 2,4 ethyl groups, does not possess the extended conjugation seen for native CN-Protohemin with its 2,4 vinyl groups. Indeed, reduced polarity of CN-Mesohemin over that of CN-Protohemin resulted in distinct temperature dependencies. Molecular visualization and protein-ligand interaction analysis targeted a functionally diverse residue unique to the α-chain. Tyrosine-42 (a polar/non-polar amino acid) appeared to play a prominent role in the assembly process.


Apohemoglobin A CN-Mesohemin protein–ligand interactions Soret region