TY - JOUR
T1 - Identification of proteins from human permanent erupted enamel
AU - Castiblanco, Gina A.
AU - Rutishauser, Dorothea
AU - Ilag, Leopold L.
AU - Martignon, Stefania
AU - Castellanos, Jaime E.
AU - Mejía, Wilson
N1 - Publisher Copyright:
© 2015 European Journal of Oral Sciences.
PY - 2015/12/1
Y1 - 2015/12/1
N2 - 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.
AB - 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.
KW - Amelogenin
KW - Dental enamel proteins
KW - Enamelin
KW - Mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=84947918109&partnerID=8YFLogxK
U2 - 10.1111/eos.12214
DO - 10.1111/eos.12214
M3 - Article
C2 - 26432388
AN - SCOPUS:84947918109
SN - 0909-8836
VL - 123
SP - 390
EP - 395
JO - European Journal of Oral Sciences
JF - European Journal of Oral Sciences
IS - 6
ER -