Coelenterazine sulfotransferase from Renilla muelleri

PLoS One. 2022 Oct 17;17(10):e0276315. doi: 10.1371/journal.pone.0276315. eCollection 2022.

Abstract

The luciferin sulfokinase (coelenterazine sulfotransferase) of Renilla was previously reported to activate the storage form, luciferyl sulfate (coelenterazine sulfate) to luciferin (coelenterazine), the substrate for the luciferase bioluminescence reaction. The gene coding for the coelenterazine sulfotransferase has not been identified. Here we used a combined proteomic/transcriptomic approach to identify and clone the sulfotransferase cDNA. Multiple isoforms of coelenterazine sulfotransferase were identified from the anthozoan Renilla muelleri by intersecting its transcriptome with the LC-MS/MS derived peptide sequences of coelenterazine sulfotransferase purified from Renilla. Two of the isoforms were expressed in E. coli, purified, and partially characterized. The encoded enzymes display sulfotransferase activity that is comparable to that of the native sulfotransferase isolated from Renilla reniformis that was reported in 1970. The bioluminescent assay for sensitive detection of 3'-phosphoadenosine 5'-phosphate (PAP) using the recombinant sulfotransferase is demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arylsulfotransferase
  • Chromatography, Liquid
  • DNA, Complementary
  • Escherichia coli* / genetics
  • Imidazoles
  • Luciferases / genetics
  • Luminescent Measurements
  • Proteomics*
  • Pyrazines
  • Renilla / genetics
  • Sulfates
  • Sulfotransferases / genetics
  • Tandem Mass Spectrometry

Substances

  • DNA, Complementary
  • Imidazoles
  • Pyrazines
  • Sulfates
  • coelenterazine
  • Luciferases
  • Sulfotransferases
  • Arylsulfotransferase

Grants and funding

This work was funded internally by New England Biolabs, Ipswich, MA, of which all authors are employees. The funder provided support in the form of salaries for: G.T., B.B., J.B., E.B., I.R.C., L.E., C.M., and I.S., but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.