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Prolonged heat stress of Lactobacillus paracasei GCRL163 improves binding to human colorectal adenocarcinoma HT-29 cells and modulates the relative abundance of secreted and cell surface-located proteins

Citation

Adu, KT and Wilson, R and Baker, AL and Bowman, J and Britz, ML, Prolonged heat stress of Lactobacillus paracasei GCRL163 improves binding to human colorectal adenocarcinoma HT-29 cells and modulates the relative abundance of secreted and cell surface-located proteins, Journal of Proteome Research, 19, (4) pp. 1824-1846. ISSN 1535-3893 (2020) [Refereed Article]

Copyright Statement

Copyright 2020 American Chemical Society

DOI: doi:10.1021/acs.jproteome.0c00107

Abstract

Lactobacillus casei group bacteria improve cheese ripening and may interact with host intestinal cells as probiotics, where surface proteins play a key role. Three complementary methods [trypsin shaving (TS), LiCl–sucrose (LS) extraction, and extracellular culture fluid precipitation] were used to analyze cell surface proteins of Lactobacillus paracasei GCRL163 by label-free quantitative proteomics after culture to the mid-exponential phase in bioreactors at pH 6.5 and temperatures of 30–45 °C. A total of 416 proteins, including 300 with transmembrane, cell wall anchoring, and secretory motifs and 116 cytoplasmic proteins, were quantified as surface proteins. Although LS caused significantly greater cell lysis as growth temperature increased, higher numbers of extracytoplasmic proteins were exclusively obtained by LS treatment. Together with the increased positive surface charge of cells cultured at supra-optimal temperatures, proteins including cell wall hydrolases Msp1/p75 and Msp2/p40, α-fucosidase AlfB, SecA, and a PspC-domain putative adhesin were upregulated in surface or secreted protein fractions, suggesting that cell adhesion may be altered. Prolonged heat stress (PHS) increased binding of L. paracasei GCRL163 to human colorectal adenocarcinoma HT-29 cells, relative to acid-stressed cells. This study demonstrates that PHS influences cell adhesion and relative abundance of proteins located on the surface, which may impact probiotic functionality, and the detected novel surface proteins likely linked to the cell cycle and envelope stress.

Item Details

Item Type:Refereed Article
Keywords:cell surface proteins, trypsin shaving, LiCl, proteomics, heat stress, cell lysis, cell adhesion, hydrophobicity, Lactobacillus casei/paracasei, cell wall hydrolases, alpha-fucosidase, PspC
Research Division:Biological Sciences
Research Group:Biochemistry and cell biology
Research Field:Proteomics and intermolecular interactions (excl. medical proteomics)
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the biological sciences
UTAS Author:Adu, KT (Mr Kayode Adu)
UTAS Author:Wilson, R (Dr Richard Wilson)
UTAS Author:Baker, AL (Dr Anthony Baker)
UTAS Author:Bowman, J (Associate Professor John Bowman)
UTAS Author:Britz, ML (Professor Margaret Britz)
ID Code:138706
Year Published:2020
Deposited By:Central Science Laboratory
Deposited On:2020-04-23
Last Modified:2020-12-08
Downloads:0

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