TY - JOUR
T1 - Isolation and full characterisation of a potentially allergenic lipid transfer protein (LTP) in almond
AU - Buhler, Sofie
AU - Tedeschi, Tullia
AU - Faccini, Andrea
AU - Garino, Cristiano
AU - Arlorio, Marco
AU - Dossena, Arnaldo
AU - Sforza, Stefano
N1 - Publisher Copyright:
© 2015, © 2015 Taylor & Francis.
PY - 2015/5/4
Y1 - 2015/5/4
N2 - Non-specific lipid transfer proteins (nsLTP) were shown to be among the most significant allergens, in particular in several fruits belonging to the Rosaceae family. The molecular features of LTPs, such as the presence of eight cysteine residues forming four disulfide bridges, confer a compact structure, decreasing the probability of degradation due to cooking or digestion, thereby increasing the chance of systemic absorption and severe allergic reactions. Few studies on LTP-induced allergies regarding almond (Prunus dulcis L) are available in the literature. In the present work, we describe for the first time the extraction and purification of an almond LTP, achieving its full characterisation by using liquid chromatography and exact mass spectrometry; the full sequence was identified by means of LC-ESI-Orbitrap™-MS applying a bottom-up approach. The characterised protein consists of 92 amino acids and has a calculated exact MW of 9579.0. The presence of four disulfide bridges was confirmed after reduction, as shown by a mass increment of 8 Da. Finally, its potential allergenicity was confirmed via an in silico approach. The results presented here demonstrate the enormous potential of advanced MS techniques for obtaining high-quality structural and functional data of allergenic proteins in a short time.
AB - Non-specific lipid transfer proteins (nsLTP) were shown to be among the most significant allergens, in particular in several fruits belonging to the Rosaceae family. The molecular features of LTPs, such as the presence of eight cysteine residues forming four disulfide bridges, confer a compact structure, decreasing the probability of degradation due to cooking or digestion, thereby increasing the chance of systemic absorption and severe allergic reactions. Few studies on LTP-induced allergies regarding almond (Prunus dulcis L) are available in the literature. In the present work, we describe for the first time the extraction and purification of an almond LTP, achieving its full characterisation by using liquid chromatography and exact mass spectrometry; the full sequence was identified by means of LC-ESI-Orbitrap™-MS applying a bottom-up approach. The characterised protein consists of 92 amino acids and has a calculated exact MW of 9579.0. The presence of four disulfide bridges was confirmed after reduction, as shown by a mass increment of 8 Da. Finally, its potential allergenicity was confirmed via an in silico approach. The results presented here demonstrate the enormous potential of advanced MS techniques for obtaining high-quality structural and functional data of allergenic proteins in a short time.
KW - LTP
KW - LTQ-Orbitrap
KW - almonds
KW - food allergy
KW - proteomics
UR - http://www.scopus.com/inward/record.url?scp=84961289128&partnerID=8YFLogxK
U2 - 10.1080/19440049.2015.1016123
DO - 10.1080/19440049.2015.1016123
M3 - Article
SN - 1944-0049
VL - 32
SP - 648
EP - 656
JO - Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment
JF - Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment
IS - 5
ER -