4605
Accession Number
31345
Author
Solazzo, Caroline;Fitzhugh, William W.;Rolando, Christian;Tokarski, Caroline
Title Of Article Chaper
Identification of Protein Remains in Archaeological Potsherds by Proteomics
Title Of Journal Book
Analytical Chemistry
Volume
80
Issue
12
Pages
4590-4597
Collation
8 p. : ills.
Reference Bibliography
Includes bibliographic references
Language Of Text
English
Literature Type
Serial
Literature Level
Analytic
Abstract
We demonstrate here the possibility of identifying proteins trapped in few milligrams of the clay matrix of a 1200-1400 AD Inupiat potsherd fragment from Point Barrow, Alaska, by a dedicated proteomics approach. The four main steps of a proteomics analysis, (i) protein extraction from biological samples, (ii) protein hydrolysis using a hydrolase enzyme, (iii) nanoLC, nanoESI MS, and MS/MS analysis of the generated peptides, and (iv) protein identification using protein databank proceeded from genomic data, have been optimized for archaeological remains. Briefly our procedures states by grinding the potsherds, extraction with 1% trifluoro-acetic acid, digestion with excess of trypsin, nanoLC, nanoESI FT-ICR analysis, and data mining by homology search. The developed conditions were evaluated on protein extracts from remains obtained by heated muscle tissues and blubbers of different seal and whale species, these samples representing the main diet sources of the Eskimo population. Most of the proteins were identified by sequence homology to other species due to the lack of cetacean and pinniped proteins in the databanks. More interestingly, two proteins, myoglobin and hemoglobin, respectively, identified in muscle tissue samples and blubber samples highlight several specific peptides of cetacean and pinniped species; these peptides are significant to prove the presence of these marine species in the analyzed samples. Based on the developed methodology and on protein identification results obtained from the heated seal/whale muscle tissues and blubbers, the analysis of the clay matrix of a 1200-1400 AD Inupiat potsherd fragment from Point Barrow was investigated. The described method succeeds in identifying for peptides corresponding to the harbor seal myoglobin (species Phoca vitulina) with a measured mass accuracy better than 1 ppm (MS and MS/MS experiments) including one specific peptide of the cetacean and pinniped species and on specific peptide of the seal species. These results highlight, for the first time, a methodology able to identify proteins from a few milligrams of archaeological potsherd buried for years; the obtained results confirm the presence of a seal muscle tissue protein in this Punuk potsherd.
Keywords
identification;protein;remain;archaeological;pot;potsherd;sherd;proteomics
pub_id
4605