Conformational and functional significance of the α140 side-chain in HbA: A UV and visible resonance Raman study of three α140 mutants

Laura Juszczak, Uri Samuni, Joel M. Friedman

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Continuous wave UV resonance Raman (UVRR) and nanosecond time-resolved visible resonance Raman spectra were generated from the deoxy and CO derivatives of three α-chain mutants of human adult hemoglobin (HbA): Yα140F, Yα140G and Yα140A. The UVRR results show that an aromatic side-chain in the α140 site is needed for achieving the wild-type tertiary conformation in the allosterically important α 1β2 interface for both the deoxy T state and the liganded R state. The wavenumbers of ν(Fe-His), the iron-proximal histidine stretching mode, seen in the VRR spectra from the deoxy derivatives show that the hydrogen bond between α140 tyrosine and the carbonyl of α93 valine is necessary to obtain the full quaternary constraint associated with the deoxy T state even when the α1β2 interface is seemingly intact. A model is presented that describes the relationship between the packing of a cluster of residues linked to both Wβ37 and Yα140 and the extent of proximal strain/quaternary constraint at the α-chain heme. The visible resonance Raman (VRR) spectra of the 8 ns photoproduct of the CO derivatives of the three mutants in the presence and absence of allosteric effectors show that all three mutants can access the R state but they show enhanced sensitivity to added IHP. The CO derivative of the Hb(Yα140F) mutant in the presence of IHP exhibits properties consistent with the high-affinity T state observed both for non-equilibrium populations of HbA isolated using sol-gel matrices and for β37 mutants where the hinge region of the α1β2 interface is disrupted.

Original languageEnglish (US)
Pages (from-to)350-358
Number of pages9
JournalJournal of Raman Spectroscopy
Issue number4
StatePublished - Apr 2005


  • Hemoglobin
  • UV resonance Raman spectroscopy
  • α140 mutant

ASJC Scopus subject areas

  • General Materials Science
  • Spectroscopy


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