Analysis of human serum albumin variants by mass spectrometric procedures

Angela Amoresano, Annapaola Andolfo, Rosa Anna Siciliano, Rosaria Cozzolino, Lorenzo Minchiotti, Monica Galliano, Piero Pucci

Research output: Contribution to journalArticle

Abstract

A new strategy for the structural characterisation of human albumin variants has been developed which makes extensive use of mass spectrometric methodologies. The rationale behind the method is to provide a rapid and effective screening of the entire albumin structure. The first step in this strategy consists in the attempt to determine the accurate molecular mass of the intact variant by electrospray mass spectrometry often providing a first indication on the presence of the variant. An HPLC procedure has been developed to isolate all the seven fragments generated by CNBr hydrolysis of HSA in a single chromatographic step. A rapid screening of the entire albumin structure is achieved by the ESMS analysis of the peptide fragments and the protein region(s) carrying the structural abnormality is identified by its anomalous mass value(s). Mass mapping of the corresponding CNBr peptide, either by Fast Atom Bombardment Mass Spectrometry (FABMS) or by Matrix Assisted Laser Desorption Ionisation Mass Spectrometry (MALDIMS), leads to the definition of the site and the nature of the variation. This combined strategy was applied to the structural characterisation of three HSA genetic variants and provided to be an effective procedure for the rapid assessment of their structural modifications showing considerable advantages over the classical approach.

Original languageEnglish
Pages (from-to)79-92
Number of pages14
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1384
Issue number1
DOIs
Publication statusPublished - Apr 23 1998

Keywords

  • Electrospray mass spectrometry
  • Genetic variant
  • Human serum albumin
  • MALDI mass spectrometry
  • Peptide mapping

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology
  • Biophysics

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