Tyrosine B10 and heme-ligand interactions of Lucina pectinata hemoglobin II: Control of heme reactivity

Ruth Pietri, Laura Granell, Anthony Cruz, Walleska De Jesús, Ariel Lewis, Ruth Leon, Carmen L. Cadilla, Juan López Garriga

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

The distal pocket of hemoglobin II (HbII) from Lucina pectinata is characterized by the presence of a GlnE7 and a TyrB10. To elucidate the functional properties of HbII, biophysical studies were conducted on HbII and a HbI PheB10Tyr site-directed mutant. The pH titration data at neutral conditions showed visible bands at 486, 541, 577 and 605 nm for both proteins. This suggests the possible existence of a conformational equilibrium between an open and closed configuration due to the interactions of the TyrB10, ligand, and heme iron. The kinetic behavior for the reaction of both ferric proteins with H 2O2 indicates that the rate for the formation of the ferryl intermediates species varies with pH, suggesting that the reaction is strongly dependent on the conformational states. At basic pH values, the barrier for the reaction increases as the tyrosine adopts a closed conformation and the ferric hydroxyl replaces the met-aquo species. The existence of these conformers is further supported by resonance Raman (RR) data, which indicate that in a neutral environment, the ferric HbII species is present as a possible mixture of coordination and spin states, with values at 1558 and 1580 cm -1 for the ν2 marker, and 1479, 1492, and 1503 cm -1 for the ν3 mode. Moreover, the presence of the A3 and Ao conformers at 1924 and 1964 cm-1 in the HbII-CO infrared spectra confirms the existence of an open and closed conformation due to the orientation of the TyrB10 with respect to the heme active center.

Original languageEnglish (US)
Pages (from-to)195-203
Number of pages9
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1747
Issue number2
DOIs
StatePublished - Mar 14 2005

Keywords

  • Closed conformation
  • HbI PheB10Tyr mutant
  • Hemoglobin II
  • Lucina pectinata
  • Open conformation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

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