Premium
P75
Molecular screening for skin sensitisation hazard in vitro using proteomics techniques
Author(s) -
Divkovic Maja,
Basketter D,
Pease C,
Panico M,
Morris H,
Dell A
Publication year - 2004
Publication title -
contact dermatitis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.524
H-Index - 96
eISSN - 1600-0536
pISSN - 0105-1873
DOI - 10.1111/j.0105-1873.2004.00309he.x
Subject(s) - chemistry , chromatography , human serum albumin , tandem mass spectrometry , trypsin , cysteine , high performance liquid chromatography , histidine , mass spectrometry , adduct , biochemistry , amino acid , enzyme , organic chemistry
Covalent binding of hapten to skin protein is required for sensitisation. It has long been postulated that only sensitising chemicals will modify skin protein(s), whereas non‐sensitisers and irritants do not. To pinpoint the amino acid residues involved in such modifications, we employed mass spectrometry (MS) based proteomics techniques. Known sensitisers, non‐sensitisers and irritants were incubated with human serum albumin (HSA). Modified HSA samples were digested by trypsin and matrix assisted laser desorption/ionisation (MALDI‐MS) analyses were conducted on the digests. Nano‐electrospray tandem mass spectrometry (ES‐MS/MS) analyses were conducted on modified peptides purified by high performance liquid chromatography (HPLC). The HSA sequence was mapped and modified peptides identified. In HSA samples incubated with non‐sensitisers and irritants there was no change compared to HSA incubated without chemical. Modified peptides were purified by HPLC and sequenced using ES‐MS/MS. Lysine, histidine and cysteine were found to be involved in covalent modifications by the sensitising chemicals used. A double adduct of one sensitiser (MCI) on His 338 of HSA molecule was also identified. More sensitisers need to be investigated to establish similarities between covalent protein modifications of chemically related compounds. The aim is to develop an assay for protein binding ability of chemicals which could form part of a battery of tests needed to replace animal testing for sensitisation potential.