| Title: | "Identification of critical residues for the catalytic activity of ComQ, a Bacillus prenylation enzyme for quorum sensing, by using a simple bioassay system" |
| Author(s): | Hirooka K; Shioda S; Okada M; |
| Address: | "Department of Biotechnology, Faculty of Life Science and Biotechnology, Fukuyama University, Fukuyama, Hiroshima, Japan. Department of Material and Life Chemistry, Kanagawa University, Yokohama, Japan" |
| Journal Title: | Biosci Biotechnol Biochem |
| DOI: | 10.1080/09168451.2019.1685371 |
| ISSN/ISBN: | 1347-6947 (Electronic) 0916-8451 (Linking) |
| Abstract: | "Bacillus ComQ participates in the biosynthesis of a quorum-sensing signaling molecule (ComX pheromone) through catalyzing the prenylation at a Trp residue of the precursor peptide (pre-ComX) with geranyl diphosphate (C(10) type) or farnesyl diphosphate (C(15) type). We hypothesized that several residues specifically conserved among either type of ComQs are important for their substrate specificities. Using a simple bioassay, we revealed that Phe63, Asn186, and Gly190 in ComQ(RO-E-2) (C(10) type) were nondisplaceable to Ser63, Gly186, and Val190, the corresponding residues in the C(15)-type ComQ, respectively. A three-dimensional model suggested that the 186th and 190th residues are involved in the pre-ComX binding. In vitro analysis showed that substitution of Phe63 with Ser in ComQ(RO-E-2) significantly reduced the geranylation activity but substantially enhanced the farnesylation activity, whereas substitution of Ser63 with Phe in ComQ(168) (C(15) type) reduced the farnesylation activity. Therefore, the 63rd residue was found to be significant for the prenyl-substrate preference.Abbreviations: GPP: geranyl diphosphate; FPP: farnesyl diphosphate; IPP: isopentenyl diphosphate; GGPP: geranylgeranyl diphosphate; FARM: first aspartate-rich motif; SARM: second aspartate-rich motif; beta-Gal: beta-galactosidase; TBABG: tryptose blood agar base supplemented with glucose; X-gal: 5-bromo-4-chloro-3-indolyl-beta-D-galactoside" |
| Keywords: | Amino Acid Sequence Bacillus subtilis/*enzymology Bacterial Proteins/chemistry/genetics/*metabolism *Biological Assay Catalysis Diphosphates/metabolism Diterpenes/metabolism Escherichia coli/genetics Membrane Proteins/chemistry/genetics/*metabolism Mutati; |
| Notes: | "MedlineHirooka, Kazutake Shioda, Saki Okada, Masahiro eng England 2019/11/02 Biosci Biotechnol Biochem. 2020 Feb; 84(2):347-357. doi: 10.1080/09168451.2019.1685371. Epub 2019 Oct 31" |
