PDBsum entry 2pse

Oxidoreductase

PDB id: 2pse

Contents

Protein chain

305 a.a. *

Ligands

IMD

Waters ×76

* Residue conservation analysis

PDB id:

2pse

Name:

Oxidoreductase

Title:

Crystal structures of the luciferase and green fluorescent protein from renilla reniformis

Structure:

Renilla-luciferin 2-monooxygenase. Chain: a. Synonym: renilla-type luciferase. Engineered: yes. Mutation: yes

Source:

Renilla reniformis. Organism_taxid: 6136. Gene: rluc. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.50Å R-factor: 0.193 R-free: 0.232

Authors:

A.M.Loening,T.D.Fenn,S.S.Gambhir

Key ref:

A.M.Loening et al. (2007). Crystal structures of the luciferase and green fluorescent protein from Renilla reniformis. J Mol Biol, 374, 1017-1028. PubMed id: 17980388 DOI: 10.1016/j.jmb.2007.09.078

Date:

06-May-07 Release date: 22-May-07

Protein chain

P27652  (LUCI_RENRE) -  Coelenterazine h 2-monooxygenase from Renilla reniformis

Seq: 311 a.a.

Struc: 305 a.a.*

* PDB and UniProt seqs differ at 8 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.1.13.12.5 - Renilla-type luciferase.
• Pathway: Renilla-luciferin 2-monooxygenase; Watasenia-luciferin 2-monooxygenase; and Oplophorus-luciferin 2-monooxygenase
• Reaction: coelenterazine h + O2 = excited coelenteramide h monoanion + hnu + CO2 + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1016/j.jmb.2007.09.078

J Mol Biol 374:1017-1028 (2007)

PubMed id: 17980388

Crystal structures of the luciferase and green fluorescent protein from Renilla reniformis.

A.M.Loening, T.D.Fenn, S.S.Gambhir.

ABSTRACT

Due to its ability to emit light, the luciferase from Renilla reniformis (RLuc) is widely employed in molecular biology as a reporter gene in cell culture experiments and small animal imaging. To accomplish this bioluminescence, the 37-kDa enzyme catalyzes the degradation of its substrate coelenterazine in the presence of molecular oxygen, resulting in the product coelenteramide, carbon dioxide, and the desired photon of light. We successfully crystallized a stabilized variant of this important protein (RLuc8) and herein present the first structures for any coelenterazine-using luciferase. These structures are based on high-resolution data measured to 1.4 A and demonstrate a classic alpha/beta-hydrolase fold. We also present data of a coelenteramide-bound luciferase and reason that this structure represents a secondary conformational form following shift of the product out of the primary active site. During the course of this work, the structure of the luciferase's accessory green fluorescent protein (RrGFP) was also determined and shown to be highly similar to that of Aequorea victoria GFP.

Selected figure(s)

Figure 2.
Fig. 2. Structure of RrGFP. The condition used is labeled as RrGFP:PEG/MPD in Table 1. Residues 7–226 (of 233 total) were identified in the data. (a) Cartoon representation of a single unit cell of the RrGFP crystal. The four protomers in each unit cell are labeled I–IV. For each protomer, the N-terminus is shown in blue and the C-terminus is shown in red. (b) Superposition of RrGFP and AvGFP. The molecule at the center of the β-barrel is the fluorophore. The primary sequences of the two GFPs are 28% identical and 50% similar. PDB code 1EMA was used for the AvGFP structure.^58 (c) Close-up cartoon representation of the RrGFP fluorophore. The gray mesh represents a σ[A]-weighted F[o]−F[c] difference map before the inclusion of the fluorophore in the model phases, contoured at 2.0 σ.

Figure 3.
Fig. 3. (a) The topology of RLuc8's α/β-hydrolase fold domain. α-Helices are shown in blue, and β-sheets are shown in red. Numbering/lettering of the sheets/helices is done with respect to the standard for α/β-hydrolases,^33 and the locations of the presumptive catalytic residues are marked. The cap domain is an excursion from the fold pattern composed of residues 146–230 in the luciferase. (b) The domains of RLuc8. Shown are the locations of the cap domain (in gray) and α/β-hydrolase fold domain (blue to red) in the context of the crystal structure. (c) Close-up stereo cartoon representation of the active site of the RLuc8:diammonium structure. The presumptive active site residues are color coded with respect to the average degree of enzymatic perturbation mutagenesis at the site yields, based on published data.^22^,^23 Mutations at green-, yellow-, and orange-colored residues were associated with <1%, 1–10%, and 10–100%, respectively, of full enzymatic activity.

The above figures are reprinted from an Open Access publication published by Elsevier: J Mol Biol (2007, 374, 1017-1028) copyright 2007.

Figures were selected by an automated process.