PEDS Advance Access first published online on January 12, 2004
This version published online on January 21, 2004
Protein Engineering Design and Selection, doi:10.1093/protein/gzh016
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
1 Department
of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA;
Present Address: Department of Biology, Instituto de Biociências,
Universidade Estadual de São Paulo (UNESP), Rio Claro, SP, Brazil;
Department of Biology, Instituto de Biociências, Universidade Estadual de
São Paulo, Rua 24 A, PoBox 199, Rio Claro, SP,
Brazil
* To whom correspondence should be addressed. E-mail: viviani{at}rc.unesp.br.
To find out the regions having major influence on the
bioluminescence spectra of railroad worm luciferases, we constructed new
chimeric luciferases switching the fragments from residues 1-219 and from
220-545 between Phrixotrix viviani (PxvGR; Keywords:
luciferases/bioluminescence/luciferases/railroad
worms/Phrixotrix
Revised November 24, 2003
Accepted November 28, 2003
Oxford University Press
Article
The influence of
the region between residues 220-344 and beyond in Phrixotrix railroadworm
luciferases green and red bioluminescence
2 Department of Biology, Instituto de
Biociências, Universidade Estadual de São Paulo (UNESP), Rio
Claro, SP, Brazil
3 Cell
Dynamics Research Group, Division of Human Life Technology, National Institute
of Advanced Industrial Science and Technology, Osaka,
Japan
Abstract 
max
=548 nm) green-light emitting luciferase and Phrixothrix hirtus (PxhRE;
max = 623 nm) red light-emitting luciferases. Emission
spectrum (max= 571 nm) and KM for luciferin in
the chimera PxRE220GR (PxhRE: 1-219; 220-545: PxvGR) suggested that
the region above residue 220 of PxvGR had a major effect on the active site.
However, switching the sequence between the residues 220-344 from PxvGR
luciferase into PxhRE (PxREGRRE) luciferase resulted in red light emission
(max = 603 nm) indicating that the region 220-344 by itself
does not determine emission spectrum. Furthermore, the sequence before residue
220 of the green emitting luciferase is incompatible for light emission with
the sequence above residue 220 of PxhRE. These results suggest that the
fragments before and after residue 220, which correspond to distinct
subdomains, may fold differently in the green and red emitting luciferases,
affecting the active site conformation.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Riahi-Madvar and S. Hosseinkhani Design and characterization of novel trypsin-resistant firefly luciferases by site-directed mutagenesis Protein Eng. Des. Sel., November 1, 2009; 22(11): 655 - 663. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Kh. Tafreshi, S. Hosseinkhani, M. Sadeghizadeh, M. Sadeghi, B. Ranjbar, and H. Naderi-Manesh The Influence of Insertion of a Critical Residue (Arg356) in Structure and Bioluminescence Spectra of Firefly Luciferase J. Biol. Chem., March 23, 2007; 282(12): 8641 - 8647. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V.R. Viviani, F.G.C. Arnoldi, B. Venkatesh, A.J.S. Neto, F.G.T. Ogawa, A.T.L. Oehlmeyer, and Y. Ohmiya Active-Site Properties of Phrixotrix Railroad Worm Green and Red Bioluminescence-Eliciting Luciferases J. Biochem., October 1, 2006; 140(4): 467 - 474. [Abstract] [Full Text] [PDF]
|
|