Proteomics

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Optimal Processing for Proteomic Genotyping of Single Human Hairs


ABSTRACT: The human hair proteome offers new insight in the field of human identification. Hair often contains DNA that has been degraded due to the process of cornification. The smaller fragments of DNA present a challenge for obtaining useful information concerning the identity of the donor. Proteomic genotyping offers an alternative approach which starts with protein, a biomolecule abundant in the hair shaft. Nonsynonymous single nucleotide polymorphisms may be detected in the sequence of these proteins, allowing the inference of SNP genotypes. Population genetics-based information from these genotypes may be used to calculate random match probability or even infer ancestry. The current challenge of this research is to optimize processing chemistry in order to maximize genetic information from a single human hair shaft. Results indicate that optimal conditions for proteomic analysis of a single human hair include 6 hrs of reduction with 100 mM dithiothreitol at room temperature, alkylation with 200 mM iodoacetamide for 45 min, and 6 hrs of digestion with two 1:50 (enzyme:protein) additions of stabilized trypsin at room temperature, with stirring incorporated into all three steps. Our final conditions using optimized temperatures and incubation times for disulfide reduction and protein digestion produced random match probabilities of up to 1 in 624 million from a single hair with a median value of 1 in 1.1 million, compared to a maximum random match probability of 1 in 1380 and a median value of 1 in 24 for the original processing method.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hair Shaft

SUBMITTER: Glendon Parker  

LAB HEAD: Glendon John Parker

PROVIDER: PXD016155 | Pride | 2020-06-10

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
QEPlus2_01042017_27_155.raw Raw
QEPlus2_01042017_29_157.raw Raw
QEPlus2_01042017_31_156.raw Raw
QEPlus2_01042017_33_158.raw Raw
QEPlus2_01042017_35_149.raw Raw
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Publications

Optimal processing for proteomic genotyping of single human hairs.

Goecker Zachary C ZC   Salemi Michelle R MR   Karim Noreen N   Phinney Brett S BS   Rice Robert H RH   Parker Glendon J GJ  

Forensic science international. Genetics 20200525


The use of hair evidence for human identification is undergoing considerable improvement through the adoption of proteomic genotyping. Unlike traditional microscopic comparisons, protein sequencing provides quantitative and empirically based estimates for random match probability. Non-synonymous SNPs are translated as single amino acid polymorphisms and result in genetically variant peptides. Using high resolution mass spectrometry, these peptides can be detected in hair shaft proteins and used  ...[more]

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