Document Type

Article

Original Publication Date

2017

Journal/Book/Conference Title

Journal of Molecular Biology Research

Volume

7

Issue

1

First Page

50

Last Page

57

DOI of Original Publication

10.5539/jmbr.v7n1p50

Date of Submission

June 2017

Abstract

Streptococcus sanguinis is a commensal and early colonizer of oral cavity as well as an opportunistic pathogen of infectious endocarditis. Extracting the soluble proteome of this bacterium provides deep insights about the physiological dynamic changes under different growth and stress conditions, thus defining “proteomic signatures” as targets for therapeutic intervention. In this protocol, we describe an experimentally verified approach to extract maximal cytoplasmic proteins from Streptococcus sanguinis SK36 strain. A combination of procedures was adopted that broke the thick cell wall barrier and minimized denaturation of the intracellular proteome, using optimized buffers and a sonication step. Extracted proteome was quantitated using Pierce BCA Protein Quantitation assay and protein bands were macroscopically assessed by Coomassie Blue staining. Finally, a high resolution detection of the extracted proteins was conducted through Synapt G2Si mass spectrometer, followed by label-free relative quantification via Progenesis QI. In conclusion, this pipeline for proteomic extraction and analysis of soluble proteins provides a fundamental tool in deciphering the biological complexity of Streptococcus sanguinis.

Is Part Of

VCU Philips Institute for Oral Health Research Publications

Recommended Citation

El-Rami, F., Nelson, K., Xu, P. (2017) Proteomic Approach for Extracting Cytoplasmic Proteins from Streptococcus sanguinis using Mass Spectrometry. Journal of Molecular Biology Research; Vol. 7, No. 1; 2017