Observation of albumin resonances in proton nuclear magnetic resonance spectra of human blood plasma : N-terminal assignments aided by use of modified recombinant albumin

Two-dimensional total shift correlation spectroscopy (TOCSY) and double-quantum-filtered phase-sensitive homonuclear shift-correlated spectroscopy (DQF-COSY) 1H NMR spectra are used to assign peaks for about one sixth of the amino acids residues of isolated human serum albumin (67 kDa) to amino acid...

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Published inAnalyst (London) Vol. 121; no. 7; pp. 913 - 922
Main Authors HARRIS, R, PATEL, S. U, SADLER, P. J, VILES, J. H
Format Journal Article
LanguageEnglish
Published Cambridge Royal Society of Chemistry 01.07.1996
Subjects
Amino Acid Sequence
Analysis
Biological and medical sciences
General pharmacology
Humans
Lipoproteins - blood
Magnetic Resonance Spectroscopy
Medical sciences
Molecular Sequence Data
Pharmacology. Drug treatments
Recombinant Proteins - chemistry
Serum Albumin - chemistry
Human
Drug
Biological fluid
Biological fixation
Albumin
NMR spectrometry
Measurement method
Recombinant protein
Blood plasma
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Summary:Two-dimensional total shift correlation spectroscopy (TOCSY) and double-quantum-filtered phase-sensitive homonuclear shift-correlated spectroscopy (DQF-COSY) 1H NMR spectra are used to assign peaks for about one sixth of the amino acids residues of isolated human serum albumin (67 kDa) to amino acid types. Sequential assignments are presented for 1H NMR resonances of the N-terminal residues Asp1, Ala2 and His3 of human serum albumin (HSA). These are based on pH-dependent chemical shifts reflecting the titrating N-terminal NH2 and the His3 imidazole ring, in addition to DQF-COSY and TOCSY experiments. Studies of variant recombinant human albumin with Asp1 deleted, rHA(2-585), aided the assignments. The structural nature of the N-and C-termini of HSA are discussed and pKa values of 7.9 and 6.3 were determined for the N-terminal amino group and His3 imidazole ring, respectively. About 20 spin systems for albumin, including those for the N-terminal amino acids, were assigned in 1H NMR spectra of blood plasma buy comparison with isolated albumin. Resonances for lipids within lipoproteins and also several low molecular mass components can also be assigned in 2D TOCSY 1H NMR spectra of plasma.
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ISSN:0003-2654
1364-5528
DOI:10.1039/AN9962100913