7VSX
Crystal structure of QL-nanoKAZ (Reverse mutant of nanoKAZ with L18Q and V27L)
Summary for 7VSX
| Entry DOI | 10.2210/pdb7vsx/pdb |
| Descriptor | QLnK, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID (3 entities in total) |
| Functional Keywords | mutated 19 kda protein, oplophorus luciferase, luminescent protein |
| Biological source | Oplophorus gracilirostris |
| Total number of polymer chains | 1 |
| Total formula weight | 19383.09 |
| Authors | Tomabechi, Y.,Sekine, S.,Shirouzu, M.,Takamitsu, H.,Satoshi, I. (deposition date: 2021-10-27, release date: 2022-08-24, Last modification date: 2023-11-29) |
| Primary citation | Inouye, S.,Sato, J.I.,Sahara-Miura, Y.,Tomabechi, Y.,Sumida, Y.,Sekine, S.I.,Shirouzu, M.,Hosoya, T. Reverse mutants of the catalytic 19 kDa mutant protein (nanoKAZ/nanoLuc) from Oplophorus luciferase with coelenterazine as preferred substrate. Plos One, 17:e0272992-e0272992, 2022 Cited by PubMed Abstract: Native Oplophorus luciferase (OpLase) and its catalytic 19 kDa protein (wild KAZ) show highest luminescence activity with coelenterazine (CTZ) among CTZ analogs. Mutated wild KAZ with 16 amino acid substitutions (nanoKAZ/nanoLuc) utilizes bis-coelenterazine (bis-CTZ) as the preferred substrate and exhibits over 10-fold higher maximum intensity than CTZ. To understand the substrate selectivity of nanoKAZ between CTZ and bis-CTZ, we prepared the reverse mutants of nanoKAZ by amino acid replacements with the original amino acid residue of wild KAZ. The reverse mutant with L18Q and V27L substitutions (QL-nanoKAZ) exhibited 2.6-fold higher maximum intensity with CTZ than that of nanoKAZ with bis-CTZ. The catalytic properties of QL-nanoKAZ including substrate specificity, luminescence spectrum, luminescence kinetics, luminescence products of CTZ, and luminescence inhibition by deaza-CTZ analogs were characterized and were compared with other CTZ-utilizing luciferases such as Gaussia and Renilla luciferases. Thus, QL-nanoKAZ with CTZ could be used as a potential reporter protein for various luminescence assay systems. Furthermore, the crystal structure of QL-nanoKAZ was determined at 1.70 Å resolution. The reverse mutation at the L18Q and V27L positions of α2-helix in nanoKAZ led to changes in the local structures of the α4-helix and the β6- and β7-sheets, and might enhance its binding affinity and oxidation efficiency with CTZ to emit light. PubMed: 36129943DOI: 10.1371/journal.pone.0272992 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.698 Å) |
Structure validation
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