Protein extraction from human anagen head hairs 1-millimeter or less in total length

Abstract

A simple method for extracting protein from human anagen (i.e., actively growing hair stage) head hairs was developed in this study for cases of limited sample availability and/or studies of specific micro-features within a hair. The distinct feature segments of the hair from one donor were divided lengthwise (i.e., each of ∼200–400 μm) and then pooled for three individual hairs to form a total of eight composite hair samples (i.e., each of ∼1 mm or less in total length). The proteins were extracted, digested using trypsin, and characterized via nano-flow liquid chromatography tandem-mass spectrometry (nLC-MS/MS). A total of 63 proteins were identified from all eight protein samples analyzed of which 60% were keratin and keratin-associated proteins. The major hair keratins identified are consistent with previous studies using fluorescence in situ hybridization and nLC-MS/MS while requiring over 400–8000-fold less sample. The protein extraction method from micro-sized human head hairs described in this study will enable proteomic analysis of biological evidence for cases of limited sample availability and will complement hair research. For example, research seeking to develop alternative non-DNA based techniques for comparing questioned to known hairs, and understanding the biochemistry of hair decomposition.

A Combined Multi-Cohort Approach Reveals Novel and Known Genome-Wide Selection Signatures for Wool Traits in Merino and Merino-Derived Sheep Breeds

“…LOC101120019 —A mutation in a 60S ribosomal protein (L21) has been shown to be involved in hereditary hypotrichosis simplex (HHS), a form of nonsyndromic inherited hair loss disorders (Zhou et al, 2011). 60S ribosomal proteins (L6 and L24) have been shown to be expressed in human anagen hair samples (Carlson et al, 2018). Interestingly, the 60S ribosomal protein L10A has been shown to be expressed in root hairs of Medicago truncatula (Covitz et al, 1998).…”

Development of a Protein‐based Human Identification Capability from a Single Hair

“…Due to extensive disulfide and isopeptide bonds, hair is an extremely difficult matrix to fully dissolve . Human hair samples of 1 mm length have been analyzed in a previous study aimed at identifying the major proteins observed in such small sample sizes . In the present study, a combination of surfactants, elevated temperatures, and ultrasonication was employed to achieve complete dissolution of single 1‐inch (25 mm) hair segments to identify potential GVPs.…”

Sensitive Method for the Confident Identification of Genetically Variant Peptides in Human Hair Keratin

“…A recent publication by Mason et al described protein‐based or GVP‐based human identification from a single hair as short as 1 inch long. Another recent publication by Carlson et al described a sensitive method to extract proteins from 1 millimeter or less in total length of human anagen head hairs and compared the proteins identified from hair shaft and hair root. The effectiveness of this method for detecting GVPs has not yet been determined.…”