Dataset Information

SigB modulates expression of novel SigB regulon members via Bc1009 in non-stressed and heat-stressed cells revealing its alternative roles in Bacillus cereus

ABSTRACT: Protein samples were measured using the LC-MS/MS platform containing reversed-phase nano liquid chromatography (nano Acquity M-class UPLC) (Waters Corporation, Massachusetts, USA), which was coupled to nanospray ionization tandem mass spectrometry with traveling wave ion mobility using high-definition data-independent (HD-MSE) acquisition and enabled with hybrid quadrupole orthogonal acceleration time of flight mass spectrometer (Synapt G2Si, Waters Corporation). The peptide mixture was separated on ACQUITY UPLC M-Class HSS T3 1.8 um, 75 um x 200 mm column (Waters Corporation) using a mixture of Buffers A (0.1% v/v Acetic acid in water) and B (0.1% v/v acetic acid in acetonitrile) through a linear concentration gradient of Buffer B in 170 min, with a flow rate of 300 nl/min from 5-26% (v/v). The eluents were sprayed at a voltage of 1.85-1.90 kV using PicoTip emitters (Waters Corporation), and other source parameters were set (sampling cone = 40V, source offset = 0 V, source temperature = 80C, gas flows, cone gas = 50 l/h, nano gas flow = 0.4 bar, purge gas = unchanged, IMS = optimized for wave velocity with a start velocity of 870 m/s to end at 564 m/s that corresponds to the separation of GluFib fragments in the drift time range of 0-200 bins). The data was acquired using the MassLynx software program version V4.1 (Waters Corporation) that automatically switches between MS and MS/MS (HDMSE) scans in resolution mode at 20000 with 1 second (sec) scan time and the scan range of 50-2000 m/z. Calibration was done by injecting GluFib at an interval of 1 min. The acquired data were analyzed for protein identifications against the Uniprot fasta database of B. cereus ATCC14579 strain that contained 5240 sequences via the PLGS v3.3 program (Water Corporation). Spectra were processed using low and high-energy thresholds of 135, 20 counts, and lock mass calibration 785.8456 m/z for GluFib. The workflow search parameters were set as: (protease = trypsin, one missed cleavage, carbamidomethyl for cysteine as fixed, variable modifier = oxidation of methionine). The protein was quantified when 1) the top 3 peptides had no modifications; 2) pass one match had peptide fragment one; and 3) ranked the first three highest peptides. The PLGS independent identification output was imported to ISOQuant 1.8 (Distler et al., 2014) for sample analyses. [doi:10.25345/C5Z892K4R] [dataset license: CC0 1.0 Universal (CC0 1.0)]

INSTRUMENT(S): SYNAPT G2-Si

ORGANISM(S): Bacillus Cereus Atcc 14579 (ncbitaxon:226900)

SUBMITTER: Prof. Dr. Uwe Voelker

PROVIDER: MSV000090315 | MassIVE |

REPOSITORIES: MassIVE

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SigB modulates expression of novel SigB regulon members via Bc1009 in non-stressed and heat-stressed cells revealing its alternative roles in Bacillus cereus.

Yeak Kah Yen Claire KYC Tempelaars Marcel M Wu Jia Lun JL Westerveld Wouter W Reder Alexander A Michalik Stephan S Dhople Vishnu M VM Völker Uwe U Pané-Farré Jan J Wells-Bennik Marjon H J MHJ Abee Tjakko T

BMC microbiology 20230210 1

<h4>Background</h4>The Bacillus cereus Sigma B (SigB) dependent general stress response is activated via the two-component RsbKY system, which involves a phosphate transfer from RsbK to RsbY. It has been hypothesized that the Hpr-like phosphocarrier protein (Bc1009) encoded by bc1009 in the SigB gene cluster may play a role in this transfer, thereby acting as a regulator of SigB activation. Alternatively, Bc1009 may be involved in the activation of a subset of SigB regulon members.<h4>Results</h ...[more]

PMID: 36759782

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