María D Contreras-Aguilar1, Jerome Vialaret2, Dominique Deville de Périère2, Damián Escribano3, Sylvain Lehmann2, Fernando Tecles1, Jose J Cerón4, Christophe Hirtz2. 1. Clinic Analysis Interdisciplinary Laboratory (Interlab-UMU), Campus of Excellence Mare Nostrum, Murcia, Spain. 2. LBPC/PPC - IRMB, CHU de Montpellier, INSERM, Montpellier University, 80 rue Augustin Fliche, Montpellier, France. 3. Department of Animal and Food Science, Veterinary School, Universitat Autònoma de Barcelona, Barcelona, Spain. 4. Clinic Analysis Interdisciplinary Laboratory (Interlab-UMU), Campus of Excellence Mare Nostrum, Murcia, Spain. jjceron@um.es.
Abstract
OBJECTIVES: To evaluate the sAA proteoforms' expression during different stimulation situations. MATERIALS AND METHODS: This study evaluated the salivary alpha-amylase (sAA) proteoforms' behavior by western blot (WB) analysis and high-resolution mass spectrometry (LC-MS/MS) in different situations that produce increases in sAA activity. For this purpose, six healthy women with a similar body mass index, age, and fit, underwent different sAA stimulation tests, such as acetic acid stimulation, psychological stress using the standardized Trier social stress test, and physical effort using the Cooper treadmill test. RESULTS: The three models showed an increase in sAA activity. The WB demonstrated seven common bands observed in the six women (band one at 59 kDa, two at 56 kDa, three at 48 kDa, four at 45 kDa, five at 41 kDa, six at 36 kDa, and seven at 14 kDa), in which sAA protein was identified. The individual WB analysis showed that band two, which corresponded to the native non-glycosylated sAA proteoform, had a higher increase after the three sAA stimulation inducers, and this band was also the only proteoform correlated with sAA activity (r = 0.56, P = 0.001). In addition, when the label-free quantification analysis was performed, the different proteoforms showed different responses depending on the type of stimulation. CONCLUSIONS: This preliminary study showed that the diverse sAA proteoforms' expression depends on the different stimulation models. CLINICAL RELEVANCE: This study opens new perspectives and challenges for the use of the different alpha-amylase proteoforms as possible biomarkers in addition to the sAA activity.
OBJECTIVES: To evaluate the sAA proteoforms' expression during different stimulation situations. MATERIALS AND METHODS: This study evaluated the salivary alpha-amylase (sAA) proteoforms' behavior by western blot (WB) analysis and high-resolution mass spectrometry (LC-MS/MS) in different situations that produce increases in sAA activity. For this purpose, six healthy women with a similar body mass index, age, and fit, underwent different sAA stimulation tests, such as acetic acid stimulation, psychological stress using the standardized Trier social stress test, and physical effort using the Cooper treadmill test. RESULTS: The three models showed an increase in sAA activity. The WB demonstrated seven common bands observed in the six women (band one at 59 kDa, two at 56 kDa, three at 48 kDa, four at 45 kDa, five at 41 kDa, six at 36 kDa, and seven at 14 kDa), in which sAA protein was identified. The individual WB analysis showed that band two, which corresponded to the native non-glycosylated sAA proteoform, had a higher increase after the three sAA stimulation inducers, and this band was also the only proteoform correlated with sAA activity (r = 0.56, P = 0.001). In addition, when the label-free quantification analysis was performed, the different proteoforms showed different responses depending on the type of stimulation. CONCLUSIONS: This preliminary study showed that the diverse sAA proteoforms' expression depends on the different stimulation models. CLINICAL RELEVANCE: This study opens new perspectives and challenges for the use of the different alpha-amylase proteoforms as possible biomarkers in addition to the sAA activity.
Authors: Andrew Thompson; Jürgen Schäfer; Karsten Kuhn; Stefan Kienle; Josef Schwarz; Günter Schmidt; Thomas Neumann; R Johnstone; A Karim A Mohammed; Christian Hamon Journal: Anal Chem Date: 2003-04-15 Impact factor: 6.986
Authors: Jon-Kar Zubieta; Mary M Heitzeg; Yolanda R Smith; Joshua A Bueller; Ke Xu; Yanjun Xu; Robert A Koeppe; Christian S Stohler; David Goldman Journal: Science Date: 2003-02-21 Impact factor: 47.728
Authors: George H Perry; Nathaniel J Dominy; Katrina G Claw; Arthur S Lee; Heike Fiegler; Richard Redon; John Werner; Fernando A Villanea; Joanna L Mountain; Rajeev Misra; Nigel P Carter; Charles Lee; Anne C Stone Journal: Nat Genet Date: 2007-09-09 Impact factor: 38.330
Authors: María D Contreras-Aguilar; Sandra V Mateo; Fernando Tecles; Christophe Hirtz; Damián Escribano; Jose J Cerón Journal: Biology (Basel) Date: 2021-03-16