New techniques for the recovery of small amounts of mature enamel proteins

Porto, Isabel Maria; Laure, Hélen Julie; Sousa, Frederico Barbosa de; Rosa, José César; Gerlach, Raquel Fernanda

doi:10.1016/j.jas.2011.08.030

Cited by 24 publications

(34 citation statements)

References 23 publications

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“…A previous study (16) identified an amelogenin peptide by comparison of the MS signal obtained with that of a synthetic peptide as the signal intensity was not high enough to allow the amino acid sequence to be identified directly from the MS data. Our samples gave MS peaks of sufficient resolution for direct identification of amino acids, in agreement with a recent study by our group using the micro‐etching technique in contemporary mature human enamel (13). Furthermore, the technique used in this study is more conservative, preserves the dental crown, and avoids contamination with collagen from dentine, which occurred in the previous study (16).…”

Section: Discussionsupporting

confidence: 90%

Recovery and identification of mature enamel proteins in ancient teeth

Porto

Laure

Tykot

et al. 2011

European J Oral Sciences

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Proteins in mineralized tissues provide a window to the past, and dental enamel is peculiar in being highly resistant to diagenesis and providing information on a very narrow window of time, such as the developing period; however, to date, complete proteins have not been extracted successfully from ancient teeth. In this work we tested the ability of a whole-crown micro-etch technique to obtain enamel protein samples from mature enamel of recently extracted (n = 2) and ancient (n = 2; ad 800 to 1100) third molars. Samples were analyzed using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry, and the resulting spectra were searched against the Swiss-Prot protein database using the Mascot software for protein identification. In our protocol, the separation of proteins in gel is not necessary. Successful identification of specific enamel proteins was obtained after whole-crown superficial enamel etching with 10% HCl. Most protein fragments recovered from dry teeth and mummy teeth contained amino-terminal amelogenin peptides. Only one peptide specific for the amelogenin X-isoform was identified. In conclusion, the reported techniques allowed the successful recovery of proteins specific to dental enamel from samples obtained in a very conservative manner, which may also be important in forensic and/or archeological science.

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Section: Discussionsupporting

confidence: 90%

Recovery and identification of mature enamel proteins in ancient teeth

Porto

Laure

Tykot

et al. 2011

European J Oral Sciences

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“…it may prove useful for non‐destructive enamel protein‐extraction procedures, such as those previously reported for sex identification of ancient specimens) . Our method increases the efficiency of identification of enamel‐specific proteins, reducing the occurrence of laboratory contaminants, such as keratin, reported in similar studies . This was accomplished by using the external third of enamel and by the omission of trypsin digestion.…”

Section: Discussionmentioning

confidence: 82%

“…Previous studies have used up to five molar samples for MS identifications ; in contrast, our study used eight molars and thus is the largest set of samples reported so far for the identification of enamel proteins from erupted sound human molars using MS. In this study, we sequenced a much broader set of amelogenin peptides than previously reported, and six were present in all samples (Table ).…”

Section: Discussionmentioning

confidence: 99%

Identification of proteins from human permanent erupted enamel

Castiblanco

Rutishauser

Ilag

et al. 2015

European J Oral Sciences

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Proteins from the extracellular matrix of enamel are highly specific and necessary for proper enamel formation. Most proteins are removed from the matrix by enamel proteases before complete mineralization is achieved; however, some residual protein fragments persist in the mineralized matrix of erupted enamel. So far, only amelogenin peptides obtained by traditional bottom-up proteomics have been recovered and identified in human permanent erupted enamel. In this study, we hypothesize that other enamel-specific proteins are also found in human permanent enamel, by analysing human erupted third molars. Pulverized enamel was used to extract proteins, and the protein extract was subjected directly to liquid-chromatography coupled to tandem mass spectrometry (LC-MS/MS) without a previous trypsin-digestion step. Amelogenin and non-amelogenin proteins (ameloblastin and enamelin) were succesfully identified. The sequences of the naturally occurring peptides of these proteins are reported, finding in particular that most of the peptides from the amelogenin X-isoform come from the tyrosine-rich amelogenin peptide (TRAP) and that some were identified in all specimens. In conclusion, our LC-MS/MS method without trypsin digestion increased the coverage of identification of the enamel proteome from a few amelogenin peptides to a higher number of peptides from three enamel-specific proteins.

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“…It has been used to analyze proteinaceous glue from a wooden building (109), to study the photo-degradation of terpenoid varnishes (110), and to assess the condition and penetration of polyethylene glycol in preserved ships (111). There are also potential applications to pottery residues (112) and in the analysis of proteins from dental enamel (113,114). A variation of MALDI-TOF, graphite-assisted laser desorption ionisation mass spectrometry (GALDI-TOF) has also been tested in the analysis of aged triterpenoid resins in varnishes (115).…”

Section: New Mass Spectrometric Techniquesmentioning

confidence: 99%

Organic Residues in Archaeology: The Highs and Lows of Recent Research

Steele

2013

ACS Symposium Series

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The analysis of organic residues from archaeological materials has become increasingly important to our understanding of ancient diet, trade and technology. Residues from diverse contexts have been retrieved and analysed from the remains of food, medicine and cosmetics to hafting material on stone arrowheads, pitch and tar from shipwrecks, and ancient manure from soils. Research has brought many advances in our understanding of archaeological, organic residues over the past two decades. Some have enabled very specific and detailed interpretations of materials preserved in the archaeological record. However there are still areas where we know very little, like the mechanisms at work during the formation and preservation of residues, and areas where each advance produces more questions rather than answers, as in the identification of degraded fats. This chapter will discuss some of the significant achievements in the field over the past decade and the ongoing challenges for research in this area.1341280_File000001_20903114.docPrinted 7/19/2013 2

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New techniques for the recovery of small amounts of mature enamel proteins

Cited by 24 publications

References 23 publications

Recovery and identification of mature enamel proteins in ancient teeth

Recovery and identification of mature enamel proteins in ancient teeth

Identification of proteins from human permanent erupted enamel

Organic Residues in Archaeology: The Highs and Lows of Recent Research

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