Identification of S-sulfonation and S-thiolation of a novel transthyretin Phe33Cys variant from a patient diagnosed with familial transthyretin amyloidosis.
Familial transthyretin amyloidosis (ATTR) is an autosomal dominant disorder associated with a variant form of the plasma carrier protein transthyretin (TTR). Amyloid fibrils consisting of variant TTR, wild-type TTR, and TTR fragments deposit in tissues and organs. The diagnosis of ATTR relies on the identification of pathologic TTR variants in plasma of symptomatic individuals who have biopsy proven amyloid disease. Previously, we have developed a mass spectrometry-based approach, in combination with direct DNA sequence analysis, to fully identify TTR variants. Our methodology uses immunoprecipitation to isolate TTR from serum, and electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry (MS) peptide mapping to identify TTR variants and posttranslational modifications. Unambiguous identification of the amino acid substitution is performed using tandem MS (MS/MS) analysis and confirmed by direct DNA sequence analysis. The MS and MS/MS analyses also yield information about posttranslational modifications. Using this approach, we have recently identified a novel pathologic TTR variant. This variant has an amino acid substitution (Phe --> Cys) at position 33. In addition, like the Cys10 present in the wild type and in this variant, the Cys33 residue was both S-sulfonated and S-thiolated (conjugated to cysteine, cysteinylglycine, and glutathione). These adducts may play a role in the TTR fibrillogenesis.
Lim
Amareth
A
Mass Spectrometry Resource, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118, USA.
Prokaeva
Tatiana
T
McComb
Mark E
ME
Connors
Lawreen H
LH
Skinner
Martha
M
Costello
Catherine E
CE
D000595:Amino Acid Sequence; D028226:Amyloidosis, Familial / Q000097:blood / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism*; D003545:Cysteine / Q000235:genetics; D005260:Female; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics*; D018384:Oxidative Stress; D010449:Peptide Mapping; D010649:Phenylalanine / Q000235:genetics; D011228:Prealbumin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D017422:Sequence Analysis, DNA; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011228:Prealbumin; D010649:Phenylalanine; D003545:Cysteine