1H-NMR heme resonance assignments by selective deuterations in low-spin complexes of ferric hemoglobin A

Gerd N. La Mar, Thomas Jue, Kiyoshi Nagai, Kevin M. Smith, Yasuhiko Yamamoto, Robert J. Kauten, V. Thanabal, Kevin C. Langry, Ravindra K. Pandey, Hiu Kwong Leung

Research output: Contribution to journalArticle

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Abstract

The heme methyl and vinyl α-proton signals have been assigned in low-spin ferric cyanide and azide ligated derivatives of the intact tetramer of hemoglobin A, as well as the isolated chains, by reconstituting the proteins with selectively deuterated hemins. For the hemoglobin cyanide tetramer, assignment to individual subunits was effected by forming hybrid hemoglobins possessing isotope-labeled hemins in only one type of subunit. The heme methyl contact shift pattern has 1-methyl and 5-methyl shifts furthest downfield in both chains and the individual subunits of the intact hemoglobin in both the cyanide- and azide-ligated species, which is consistent with a dominant rhombic perturbation due to the proximal His-F8 imidazole π bonding in the known structure for human adult hemoglobin. The individual chain and subunit assignments confirm that the detailed electronic/magnetic properties of the heme pocket are essentially unaltered upon assembling the R-state tetramer from the isolated subunits.

Original languageEnglish (US)
Pages (from-to)131-141
Number of pages11
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume952
Issue numberC
DOIs
StatePublished - 1988

Fingerprint

Hemoglobin A
Heme
Hemin
Azides
Nuclear magnetic resonance
Cyanides
Hemoglobin Subunits
Isotopes
Protons
Magnetic properties
Hemoglobins
Derivatives
Proton Magnetic Resonance Spectroscopy
Proteins

Keywords

  • (Selective deuteration)
  • H-
  • H-
  • Ferric hemoglobin A
  • Hemoglobin
  • Low spin complex
  • NMR

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology
  • Medicine(all)

Cite this

1H-NMR heme resonance assignments by selective deuterations in low-spin complexes of ferric hemoglobin A. / La Mar, Gerd N.; Jue, Thomas; Nagai, Kiyoshi; Smith, Kevin M.; Yamamoto, Yasuhiko; Kauten, Robert J.; Thanabal, V.; Langry, Kevin C.; Pandey, Ravindra K.; Leung, Hiu Kwong.

In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, Vol. 952, No. C, 1988, p. 131-141.

Research output: Contribution to journalArticle

La Mar, Gerd N. ; Jue, Thomas ; Nagai, Kiyoshi ; Smith, Kevin M. ; Yamamoto, Yasuhiko ; Kauten, Robert J. ; Thanabal, V. ; Langry, Kevin C. ; Pandey, Ravindra K. ; Leung, Hiu Kwong. / 1H-NMR heme resonance assignments by selective deuterations in low-spin complexes of ferric hemoglobin A. In: Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular. 1988 ; Vol. 952, No. C. pp. 131-141.
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T1 - 1H-NMR heme resonance assignments by selective deuterations in low-spin complexes of ferric hemoglobin A

AU - La Mar, Gerd N.

AU - Jue, Thomas

AU - Nagai, Kiyoshi

AU - Smith, Kevin M.

AU - Yamamoto, Yasuhiko

AU - Kauten, Robert J.

AU - Thanabal, V.

AU - Langry, Kevin C.

AU - Pandey, Ravindra K.

AU - Leung, Hiu Kwong

PY - 1988

Y1 - 1988

N2 - The heme methyl and vinyl α-proton signals have been assigned in low-spin ferric cyanide and azide ligated derivatives of the intact tetramer of hemoglobin A, as well as the isolated chains, by reconstituting the proteins with selectively deuterated hemins. For the hemoglobin cyanide tetramer, assignment to individual subunits was effected by forming hybrid hemoglobins possessing isotope-labeled hemins in only one type of subunit. The heme methyl contact shift pattern has 1-methyl and 5-methyl shifts furthest downfield in both chains and the individual subunits of the intact hemoglobin in both the cyanide- and azide-ligated species, which is consistent with a dominant rhombic perturbation due to the proximal His-F8 imidazole π bonding in the known structure for human adult hemoglobin. The individual chain and subunit assignments confirm that the detailed electronic/magnetic properties of the heme pocket are essentially unaltered upon assembling the R-state tetramer from the isolated subunits.

AB - The heme methyl and vinyl α-proton signals have been assigned in low-spin ferric cyanide and azide ligated derivatives of the intact tetramer of hemoglobin A, as well as the isolated chains, by reconstituting the proteins with selectively deuterated hemins. For the hemoglobin cyanide tetramer, assignment to individual subunits was effected by forming hybrid hemoglobins possessing isotope-labeled hemins in only one type of subunit. The heme methyl contact shift pattern has 1-methyl and 5-methyl shifts furthest downfield in both chains and the individual subunits of the intact hemoglobin in both the cyanide- and azide-ligated species, which is consistent with a dominant rhombic perturbation due to the proximal His-F8 imidazole π bonding in the known structure for human adult hemoglobin. The individual chain and subunit assignments confirm that the detailed electronic/magnetic properties of the heme pocket are essentially unaltered upon assembling the R-state tetramer from the isolated subunits.

KW - (Selective deuteration)

KW - H-

KW - H-

KW - Ferric hemoglobin A

KW - Hemoglobin

KW - Low spin complex

KW - NMR

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