A Cobalamin Activity-Based Probe Enables Microbial Cell Growth and Finds New Cobalamin-Protein Interactions across Domains
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division
- Univ. of Florida, Gainesville, FL (United States). Dept. of Microbiology and Cell Science
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division; Washington State Univ., Pullman, WA (United States). School of Chemical Engineering and Bioengineering
- Purdue Univ., West Lafayette, IN (United States). Dept. of Food Science
- Univ. of Florida, Gainesville, FL (United States). Dept. of Microbiology and Cell Science and Genetics Inst.
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, here we show that a synthetic probe mimic of B12 supports the growth of B12-auxotrophic bacteria and archaea. We demonstrate that a B12 activity-based probe (B12-ABP) is actively transported into Escherichia coli cells and converted to adenosyl-B12-ABP akin to native B12. Identification of the proteins that bind the B12-ABP in vivo in E. coli, a Rhodobacteraceae sp. and Haloferax volcanii, demonstrate the specificity for known and novel B12 protein targets. The B12-ABP also regulates the B12 dependent RNA riboswitch btuB and the transcription factor EutR. Our results demonstrate a new approach to gain knowledge about the role of B12 in microbe functions. Our approach provides a powerful nondisruptive tool to analyze B12 interactions in living cells and can be used to discover the role of B12 in diverse microbial systems.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division; USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Laboratory Directed Research and Development (LDRD) Program; National Institutes of Health (NIH)
- Grant/Contract Number:
- AC05-76RL01830; FG02-05ER15650; R01-GM57498
- OSTI ID:
- 1688417
- Report Number(s):
- PNNL-SA-127734
- Journal Information:
- Applied and Environmental Microbiology, Vol. 84, Issue 18; ISSN 0099-2240
- Publisher:
- American Society for MicrobiologyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
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