Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Determination of hexahydrophthalic anhydride adducts to human serum albumin

Kåredal, Monica LU orcid ; Lindh, Christian LU orcid and Jönsson, Bo A LU (2003) In Biomarkers 8(5). p.343-359
Abstract
Hexahydrophthalic anhydride (HHPA) is a highly sensitizing industrial chemical that is known to covalently bind to endogenous proteins. The aim of this study was to determine the binding sites of HHPA to human serum albumin (HSA). Conjugates between HSA and HHPA, at two different molar ratios, were synthesized under physiological conditions. The conjugates were digested with trypsin and Pronase E to obtain specific peptides and amino acids, which were separated by liquid chromatography (LC). Fractions containing modified peptides were detected through quantification of hydrolysable HHPA using LC coupled to a triple quadrupole mass spectrometer with electrospray ionization. Modified residues in albumin were identified by sequence analyses... (More)
Hexahydrophthalic anhydride (HHPA) is a highly sensitizing industrial chemical that is known to covalently bind to endogenous proteins. The aim of this study was to determine the binding sites of HHPA to human serum albumin (HSA). Conjugates between HSA and HHPA, at two different molar ratios, were synthesized under physiological conditions. The conjugates were digested with trypsin and Pronase E to obtain specific peptides and amino acids, which were separated by liquid chromatography (LC). Fractions containing modified peptides were detected through quantification of hydrolysable HHPA using LC coupled to a triple quadrupole mass spectrometer with electrospray ionization. Modified residues in albumin were identified by sequence analyses using nanoelectrospray quadrupole time-of-flight mass spectrometry. A total of 36 HHPA adducts were found in the HSA - HHPA conjugate with 10 times molar excess of added HHPA. In the conjugate with a molar ratio of 1: 0.1 of added HHPA, seven HHPA adducts were found bound to Lys(137) (domain IB), Lys(190), Lys(199) and Lys(212) (domain IIA), Lys(351) (domain IIB), and Lys(432) and Lys(436) (domain IIIA). Moreover, several of these adducted albumin peptides were detected in nasal lavage fluid from one volunteer exposed to HHPA. The binding sites of HHPA to HSA have been determined, thus identifying potential allergenic chemical structures. This knowledge generates the possibility of developing methods for the biological monitoring of HHPA exposure by analysing tryptic peptides including these binding sites. (Less)
Please use this url to cite or link to this publication:
author
; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
liquid chromatography/mass spectrometry, organic acid anhydrides, protein adducts
in
Biomarkers
volume
8
issue
5
pages
343 - 359
publisher
Taylor & Francis
external identifiers
  • wos:000186343100001
  • scopus:0345099248
ISSN
1366-5804
DOI
10.1080/13547500310001607836
language
English
LU publication?
yes
id
a6f54c41-6e1d-4c4e-b35a-2caead9efd13 (old id 296574)
date added to LUP
2016-04-01 11:41:19
date last changed
2022-01-26 08:44:55
@article{a6f54c41-6e1d-4c4e-b35a-2caead9efd13,
  abstract     = {{Hexahydrophthalic anhydride (HHPA) is a highly sensitizing industrial chemical that is known to covalently bind to endogenous proteins. The aim of this study was to determine the binding sites of HHPA to human serum albumin (HSA). Conjugates between HSA and HHPA, at two different molar ratios, were synthesized under physiological conditions. The conjugates were digested with trypsin and Pronase E to obtain specific peptides and amino acids, which were separated by liquid chromatography (LC). Fractions containing modified peptides were detected through quantification of hydrolysable HHPA using LC coupled to a triple quadrupole mass spectrometer with electrospray ionization. Modified residues in albumin were identified by sequence analyses using nanoelectrospray quadrupole time-of-flight mass spectrometry. A total of 36 HHPA adducts were found in the HSA - HHPA conjugate with 10 times molar excess of added HHPA. In the conjugate with a molar ratio of 1: 0.1 of added HHPA, seven HHPA adducts were found bound to Lys(137) (domain IB), Lys(190), Lys(199) and Lys(212) (domain IIA), Lys(351) (domain IIB), and Lys(432) and Lys(436) (domain IIIA). Moreover, several of these adducted albumin peptides were detected in nasal lavage fluid from one volunteer exposed to HHPA. The binding sites of HHPA to HSA have been determined, thus identifying potential allergenic chemical structures. This knowledge generates the possibility of developing methods for the biological monitoring of HHPA exposure by analysing tryptic peptides including these binding sites.}},
  author       = {{Kåredal, Monica and Lindh, Christian and Jönsson, Bo A}},
  issn         = {{1366-5804}},
  keywords     = {{liquid chromatography/mass spectrometry; organic acid anhydrides; protein adducts}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{343--359}},
  publisher    = {{Taylor & Francis}},
  series       = {{Biomarkers}},
  title        = {{Determination of hexahydrophthalic anhydride adducts to human serum albumin}},
  url          = {{http://dx.doi.org/10.1080/13547500310001607836}},
  doi          = {{10.1080/13547500310001607836}},
  volume       = {{8}},
  year         = {{2003}},
}