Extraction of Non-covalently Bound Cell Surface Proteins

Non-covalently bound cell surface proteins, including SLPs and SLAPs were extracted from NCK1909 and ΔprtX L. acidophilus NCFM strains using LiCl denaturing salt, as described previously (Johnson et al., 2013 (link)). Proteins were quantified via bicinchoninic acid assay kit (Thermo Scientific) and visualized via SDS–PAGE using precast 4–20% Precise Tris-HEPES protein gels (Thermo Scientific). Gels were stained using AcquaStain (Bulldog Bio) according to the instructions of the manufacturer.

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Johnson B.R., O’Flaherty S., Goh Y.J., Carroll I., Barrangou R, & Klaenhammer T.R. (2017). The S-layer Associated Serine Protease Homolog PrtX Impacts Cell Surface-Mediated Microbe-Host Interactions of Lactobacillus acidophilus NCFM. Frontiers in Microbiology, 8, 1185.

Publication 2017

bicinchoninic acid assay kit 4–20% Precise Tris-HEPES protein gels AcquaStain

Acidophilus Assay Bicinchoninic acid Cell surface proteins Gels Hepes Protein Salt Sds page Strains Tris

Corresponding Organization : University of North Carolina at Chapel Hill

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Variable analysis

independent variables

NCK1909 strain
Δ prtX L. acidophilus NCFM strain

dependent variables

Non-covalently bound cell surface proteins, including SLPs and SLAPs
Protein quantification via bicinchoninic acid assay
Protein visualization via SDS–PAGE

control variables

LiCl denaturing salt for protein extraction, as described previously (Johnson et al., 2013)
Precast 4–20% Precise Tris-HEPES protein gels (Thermo Scientific) for SDS-PAGE
AcquaStain (Bulldog Bio) for gel staining

controls

No positive or negative controls were explicitly mentioned.

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