Journal article
Bacillus anthracis Prolyl 4-Hydroxylase Interacts with and Modifies Elongation Factor Tu
Biochemistry (Easton), Vol.56(43), pp.5771-5785
10/31/2017
DOI: 10.1021/acs.biochem.7b00601
PMCID: PMC5735995
PMID: 28981257
Abstract
Prolyl hydroxylation is a very common post-translational modification and plays many roles in eukaryotes such as collagen stabilization, hypoxia sensing, and controlling protein transcription and translation. There is a growing body of evidence that suggests that prokaryotes contain prolyl 4-hydroxylases (P4Hs) homologous to the hypoxia-inducible factor (HIF) prolyl hydroxylase domain (PHD) enzymes that act on elongation factor Tu (EFTu) and are likely involved in the regulation of bacterial translation. Recent biochemical and structural studies with a PHD from Pseudomonas putida (PPHD) determined that it forms a complex with EFTu and hydroxylates a prolyl residue of EFTu. Moreover, while animal, plant, and viral P4Hs act on peptidyl proline, most prokaryotic P4Hs have been known to target free L-proline; the exceptions include PPHD and a P4H from Bacillus anthracis (BaP4H) that modifies collagen-like proline-rich peptides. Here we use biophysical and mass spectrometric methods to demonstrate that BaP4H recognizes full-length BaEFTu and a BaEFTu 9-mer peptide for site-specific proline hydroxylation. Using size-exclusion chromatography coupled small-angle X-ray scattering (SEC SAXS) and binding studies, we determined that BaP4H forms a 1:1 heterodimeric complex with BaEFTu. The SEC SAXS studies reveal dissociation of BaP4H dimeric subunits upon interaction with BaEFTu. While BaP4H is unusual within bacteria in that it is structurally and functionally similar to the animal PHDs and collagen P4Hs, respectively, this work provides further evidence of its promiscuous substrate recognition. It is possible that the enzyme might have evolved to hydroxylate a universally conserved protein in prokaryotes, similar to the PHDs, and implies a functional role in B. anthracis.
Details
- Title: Subtitle
- Bacillus anthracis Prolyl 4-Hydroxylase Interacts with and Modifies Elongation Factor Tu
- Creators
- Nicholas J. Schnicker - University of IowaMortezaali Razzaghi - University of IowaSanjukta Guha Thakurta - Harvard UniversitySrinivas Chakravarthy - Argonne National LaboratoryMishtu Dey - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.56(43), pp.5771-5785
- DOI
- 10.1021/acs.biochem.7b00601
- PMID
- 28981257
- PMCID
- PMC5735995
- NLM abbreviation
- Biochemistry
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Publisher
- Amer Chemical Soc
- Number of pages
- 15
- Grant note
- 1S10OD018090-01 / NIGMS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) P41 GM103622 / National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) P41GM103622 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) S10OD018090 / OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA CLP 1506181 / National Science Foundation; National Science Foundation (NSF) University of Iowa College of Liberal Arts and Sciences
- Language
- English
- Date published
- 10/31/2017
- Academic Unit
- Molecular Physiology and Biophysics; Biochemistry and Molecular Biology; Chemistry; Medicine Administration
- Record Identifier
- 9984622758702771
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