Abstract
Proteomic analysis of airway smooth muscle (ASM) is key in the understanding of the regulation mechanisms of ASM contraction. The proteomic profile of ASM was evaluated in guinea pig asthma model. After ovalbumin sensitization, guinea pigs were intermittently challenged with antigen each 10 days (n=3; each group). At third challenge, isolated ASM cells from trachea were obtained. ASM cells were fractioned and processed through one- and two-dimensional gel electrophoresis and bands with molecular weights between 15-230 kDa were examined. Mass spectrometry analysis performed by MALDI/TOF-TOF MS/MS system allowed the identification of 76 proteins. Of these, protein phosphatase 1 glycogen-associated regulatory subunit (RG1), protein FAM3A, desmin, actin-binding LIM protein-2, filamin-A, collagen type I alpha-1 chain, spastic paraplegia-7 protein, annexin-A5, tropomyosin, semaphorin-3F, testis-expressed protein-15, DNA-dependent protein kinase catalytic subunit, neuroepithelial cell-transforming gene-1 protein and calponin-1 were detected in both groups but showed an increase in asthma model. The subunit regulatory of myosin phosphatase 1 (MYPT1), the main regulator of myosin light chain phosphatase 1 (MLCP), was not detected by mass spectrometry but by western blot, and showed a decrement in asthma. The analysis of proteomic interaction network revealed an inverse relation between RG1, that is an alternative regulator of MLCP, and E3 ubiquitin ligase, a protein responsible for labeling RG1 for degradation. In conclusion, it is possible that during asthma the regulation ASM relaxation through MLCP might depend on RG1 instead MYPT1. SEP-CONACyT (# 256197)
Footnotes
Cite this article as: European Respiratory Journal 2020; 56: Suppl. 64, 914.
This abstract was presented at the 2020 ERS International Congress, in session “Respiratory viruses in the "pre COVID-19" era”.
This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).
- Copyright ©the authors 2020