The 1.5-Å resolution crystal structure of bacterial luciferase in low salt conditions

Andrew J Fisher, Thomas B. Thompson, James B. Thoden, Thomas O. Baldwin, Ivan Rayment

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

Bacterial luciferase is a flavin monooxygenase that catalyzes the oxidation of a long-chain aldehyde and releases energy in the form of visible light. A new crystal form of luciferase cloned from Vibrio harveyi has been grown under low-salt concentrations, which diffract x-rays beyond 1.5-Å resolution. The x-ray structure of bacterial luciferase has been refined to a conventional R-factor of 18.2% for all recorded synchrotron data between 30.0 and 1.50-Å resolution. Bacterial luciferase is an α-β heterodimer, and the individual subunits fold into a single domain (β/α)8 barrel. The high resolution structure reveals a non-prolyl cis peptide bond that forms between Ala74 and Ala75 in the α subunit near the putative active site. This cis peptide bond may have functional significance for creating a cavity at the active site. Bacterial luciferase employs reduced flavin as a substrate rather than a cofactor. The structure presented was determined in the absence of substrates. A comparison of the structural similarities between luciferase and a nonfluorescent flavoprotein, which is expressed in the lux operon of one genus of bioluminescent bacteria, suggests that the two proteins originated from a common ancestor. However, the flavin binding sites of the nonfluorescent protein are likely not representative of the flavin binding site on luciferase. The structure presented here will furnish a detailed molecular model for all bacterial luciferases.

Original languageEnglish (US)
Pages (from-to)21956-21968
Number of pages13
JournalJournal of Biological Chemistry
Volume271
Issue number36
DOIs
StatePublished - 1996

Fingerprint

Bacterial Luciferases
Salts
Crystal structure
Luciferases
R388
Binding Sites
Catalytic Domain
Flavoproteins
X rays
Peptides
X-Rays
Substrates
Mixed Function Oxygenases
Synchrotrons
Molecular Models
Aldehydes
Operon
Bacteria
Proteins
Oxidation

ASJC Scopus subject areas

  • Biochemistry

Cite this

The 1.5-Å resolution crystal structure of bacterial luciferase in low salt conditions. / Fisher, Andrew J; Thompson, Thomas B.; Thoden, James B.; Baldwin, Thomas O.; Rayment, Ivan.

In: Journal of Biological Chemistry, Vol. 271, No. 36, 1996, p. 21956-21968.

Research output: Contribution to journalArticle

Fisher, Andrew J ; Thompson, Thomas B. ; Thoden, James B. ; Baldwin, Thomas O. ; Rayment, Ivan. / The 1.5-Å resolution crystal structure of bacterial luciferase in low salt conditions. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 36. pp. 21956-21968.
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