The position 68(E11) side chain in myoglobin regulates ligand capture, bond formation with heme iron, and internal movement into the xenon cavities

David Dantsker, Camille Roche, Uri Samuni, George Blouin, John S. Olson, Joel M. Friedman

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

After photodissociation, ligand rebinding to myoglobin exhibits complex kinetic patterns associated with multiple first-order geminate recombination processes occurring within the protein and a simpler bimolecular phase representing second-order ligand rebinding from the solvent. A smooth transition from cryogenic-like to solution phase properties can be obtained by using a combination of sol-gel encapsulation, addition of glycerol as a bathing medium, and temperature tuning (-15 → 65 °C). This approach was applied to a series of double mutants, myoglobin CO (H64L/V68X, where X = Ala, Val, Leu, Asn, and Phe), which were designed to examine the contributions of the position 68(E11) side chain to the appearance and disappearance of internal rebinding phases in the absence of steric and polar interactions with the distal histidine. Based on the effects of viscosity, temperature, and the stereochemistry of the E11 side chain, the three major phases, B → A, C → A, and D → A, can be assigned, respectively, to ligand rebinding from the following: (i) the distal heme pocket, (ii) the xenon cavities prior to large amplitude side chain conformational relaxation, and (iii) the xenon cavities after significant conformational relaxation of the position 68(E11) side chain. The relative amplitudes of the B → A and C → A phases depend markedly on the size and shape of the E11 side chain, which regulates sterically both ligand return to the heme iron atom and ligand migration to the xenon cavities. The internal xenon cavities provide a transient docking site that allows side chain relaxations and the entry of water into the vacated distal pocket, which in turn slows ligand recombination markedly.

Original languageEnglish (US)
Pages (from-to)38740-38755
Number of pages16
JournalJournal of Biological Chemistry
Volume280
Issue number46
DOIs
StatePublished - 2005

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Xenon
Myoglobin
Heme
Iron
Ligands
Genetic Recombination
Photodissociation
Stereochemistry
Temperature
Polymethyl Methacrylate
Carbon Monoxide
Encapsulation
Histidine
Viscosity
Cryogenics
Glycerol
Sol-gels
Tuning
Gels
Atoms

ASJC Scopus subject areas

  • Biochemistry

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The position 68(E11) side chain in myoglobin regulates ligand capture, bond formation with heme iron, and internal movement into the xenon cavities. / Dantsker, David; Roche, Camille; Samuni, Uri; Blouin, George; Olson, John S.; Friedman, Joel M.

In: Journal of Biological Chemistry, Vol. 280, No. 46, 2005, p. 38740-38755.

Research output: Contribution to journalArticle

Dantsker, David ; Roche, Camille ; Samuni, Uri ; Blouin, George ; Olson, John S. ; Friedman, Joel M. / The position 68(E11) side chain in myoglobin regulates ligand capture, bond formation with heme iron, and internal movement into the xenon cavities. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 46. pp. 38740-38755.
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