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. 2021 Apr 8;20(5):e13355. doi: 10.1111/acel.13355

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© 2021 The Authors. Aging Cell published by Anatomical Society and John Wiley & Sons Ltd.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Exemplar skin biomarker candidates exhibit regional differences in peptide yield due to photoageing‐specific modifications. Proteins were segmented into 50 aa‐sized step regions with average peptide counts (PSMs; N = 7) heat mapped to each step and compared between forearm and buttock (bar graphs = average PSMs, error bars = SD). Average peptide counts corresponding to each forearm protein step were subtracted from the counts of their corresponding buttock protein step and divided by the aa sequence length of that step to reveal regional differences in peptide yield (line graphs). Multiple proteins within the dermis (a–d) and epidermis (e–h) exhibited statistically significant regional differences between forearm and buttock (* ≤0.05, ** ≤0.01, *** ≤0.001; Bonferroni‐corrected repeated measures paired ANOVA). In the dermis, one region in the N‐terminal half of collagen VI alpha‐3 had significantly lower peptide counts in buttock samples than in forearm, whereas another region in the C‐terminal half of the protein had significantly higher (a). Two N‐terminal regions within fibulin‐1 had significantly higher peptide counts in buttock than in forearm (b). Biglycan had two regions near the central portion of the protein with significantly higher peptide counts in buttock than in forearm and another region on the C‐terminal side with significantly lower (c). A central region in galectin‐7 yielded significantly lower peptides in buttock samples than in forearm (d). In epidermis, two N‐terminal sided regions in K2 (e) and one in K10 (f) had significantly higher peptide counts in buttock than in forearm whereas one region near the C‐terminal end of K2 and two near the protein centre of K10 had significantly lower counts in buttock than forearm. In desmoplakin (g) one N‐terminal region and another near the C‐terminus yielded significantly higher peptides in buttock than in forearm. Lastly, HSP70 exhibited one N‐terminal sided region with significantly lower peptide counts in buttock than in forearm and another C‐terminal region with significantly higher (h)