Determination of ferric heme-human serum albumin by 1H NMR relaxometry

J Inorg Biochem. 2003 May 1;95(1):64-7. doi: 10.1016/s0162-0134(03)00070-9.

Abstract

A new method for the accurate determination of ferric heme-human serum albumin (heme-HSA) at concentrations down to the physiological level, i.e., in the micromolar concentration range, is proposed. This method is based on the (1)H NMR relaxometric properties of heme-HSA. Actually, the binding of the paramagnetic ferric heme to the primary binding site of HSA determines a strong paramagnetic enhancement of the water (1)H NMR relaxation rate. Although a linear relationship may be seen by operating at 20 MHz on conventional electromagnets, the method here reported is improved by working at 0.02 MHz on a field-cycling instrument. This (1)H NMR relaxometric method does not suffer from the presence in serum of heme catabolites (e.g., bilirubin) that affect significantly the optical determination of ferric heme-HSA in the micromolar concentration range. Paramagnetic ferric hemoglobin contribution may be selectively quenched by cyanide binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Heme / chemistry*
  • Heme / metabolism
  • Humans
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Binding
  • Serum Albumin / chemistry*
  • Serum Albumin / metabolism

Substances

  • Serum Albumin
  • Heme