Comparison of histidine proton magnetic resonances of human carbonmonoxyhaemoglobin in different buffers

J Mol Biol. 1985 Nov 20;186(2):471-3. doi: 10.1016/0022-2836(85)90119-6.

Abstract

We have recorded the C-2 proton resonances of the histidines of carbonmonoxyhaemoglobin A and of four abnormal human HbCOs in different buffers and at different concentrations of haemoglobin. Resonance H assigned by Perutz et al. (1985) to His HC3(146) beta, is present at both pH 7.30 and pH 6.90, but somewhat broadened when recorded in 5 to 10% HbCO A in 0.1 M-bis-Tris. The broadening disappears on tenfold dilution of the Hb with bis-Tris and the resonance then stands out sharply. Resonance H is absent at both Hb concentrations in HbCO Cowtown (His HC3(146) beta----Leu). HbCO Fort de France (His CD3(45) alpha----Arg) in 0.1 M-bis-Tris of pH 6.90 has a spectrum similar to that of HbCO A. In the same buffer a resonance marked L by Russu et al. (1982) is absent from the spectrum of Hb Abbruzzo (His H21(143) beta----Arg), whereas resonance H is present. Hb Barcelona contains an additional histidine in position FG1(94) beta; in 0.1 M-bis-Tris buffer of pH 6.90 its resonance is not resolved and resonance H is either shifted or broadened. The resonances of both histidines are resolved in phosphate buffer. At pH 6.90, spectra in 0.1 M-bis-Tris buffer are similar to those previously recorded in 0.2 M-HEPES. Addition of 0.1 M-KCl produces marked changes. Replacement of bis-Tris by 0.2 M-KCl + 0.2 M-phosphate gives rise to a different and much better resolved spectrum.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Buffers
  • Carboxyhemoglobin*
  • Hemoglobins*
  • Histidine*
  • Humans
  • Hydrogen-Ion Concentration
  • Magnetic Resonance Spectroscopy
  • Protons*

Substances

  • Buffers
  • Hemoglobins
  • Protons
  • Histidine
  • Carboxyhemoglobin