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Investigating B12 binding proteins in bacteria using a B12-activity-based-probe, Hv_B12ABP addendum

ABSTRACT: Elucidating the mechanistic underpinnings of a cobalamin activity-based probe in microbial cell growth, gene regulation, and enzyme activity

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Haloferax Volcanii (halobacterium Volcanii)

SUBMITTER: Matthew Monroe

LAB HEAD: Aaron Wright

PROVIDER: PXD008595 | Pride | 2018-10-22

REPOSITORIES: Pride

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			Action	DRS
	Hv_B12ABP_1noBiotin_run2_24Aug17_Merry_17-07-02.mgf	Mgf
	Hv_B12ABP_1noBiotin_run2_24Aug17_Merry_17-07-02.pride.mgf.gz	Mgf
	Hv_B12ABP_1noBiotin_run2_24Aug17_Merry_17-07-02.raw	Raw
	Hv_B12ABP_1noBiotin_run2_24Aug17_Merry_17-07-02_msgfplus.mzid.gz	Mzid
	Hv_B12ABP_1noBiotin_run2_24Aug17_Merry_17-07-02_msgfplus.pride.mztab.gz	Mztab

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Publications

A Cobalamin Activity-Based Probe Enables Microbial Cell Growth and Finds New Cobalamin-Protein Interactions across Domains.

Rosnow Joshua J JJ Hwang Sungmin S Killinger Bryan J BJ Kim Young-Mo YM Moore Ronald J RJ Lindemann Stephen R SR Maupin-Furlow Julie A JA Wright Aaron T AT

Applied and environmental microbiology 20180831 18

Understanding the factors that regulate microbe function and microbial community assembly, function, and fitness is a grand challenge. A critical factor and an important enzyme cofactor and regulator of gene expression is cobalamin (vitamin B12). Our knowledge of the roles of vitamin B12 is limited, because technologies that enable in situ characterization of microbial metabolism and gene regulation with minimal impact on cell physiology are needed. To meet this need ...[more]

PMID: 30006406

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The vitamin B12 processing enzyme, mmachc, is essential for zebrafish survival, growth and retinal morphology

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Folate and vitamin B12 imbalance induces endoplasmic reticulum stress and cholesterol biosynthesis gene expression in human adipocytes

Project description:Rationale: Low B12 has been shown to play an important role in the prediction of metabolic risk, but its significance and mechanism in the development of adiposity and adipose tissue dysfunction is largely unknown. Objective: To investigate the role of B12 and folic acid in the development of adipocyte dysfunction. Methods and Results: Microarray analysis of human adipocytes (CHUB-S7 cell line) cultured and differentiated in customised media with varying concentrations of B12 and folic acid led to the identification of two important pathways: cholesterol synthesis and unfolded protein response (UPR). Adipocytes cultured in media with low B12 (150 pmol/L) or no B12 had increased intracellular total cholesterol, higher secreted homocysteine levels, induced UPR and reduced glucose uptake capacity compared to adipocytes cultured in normal media with higher B12. The folate concentrations had either no or little effect on the measured functions. Further analysis of these adipocytes for overall DNA methylation showed that the promoter regions of sterol regulatory element-binding transcription factor 1 (SREBF1) and low density lipoprotein receptor (LDLR) were hypomethylated in the low and no B12 conditions. The SREB proteins (SREBP1 and 2) and mRNA expressions (SREBF1 and LDLR) were also increased in the same conditions. Conclusion: The data suggest that low B12 can lead to adipocyte dysfunction by inducing excess cholesterol biosynthesis, homocysteine production and induction of UPR and overall adipocyte dysfunction. Both of these pathways and adipocyte dysfunction play a significant role in the development of cardiovascular diseases. Independent replicate samples of the human adipocyte cell line CHUB-S7 were treated with four different concentrations of B12 and folate.

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An unprecedented binding mode of vitamin B12 in the predicted transporter BtuM

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2018-10-18 | PXD010024 | Pride

Effects of vitamin B12 on the gene expression of PAO1 under anaerobic growth conditions

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BtuB TonB-dependent transporters and BtuG surface lipoproteins form stable complexes for vitamin B12 uptake in gut Bacteroides.

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