2024-03-29T12:49:34Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/618772022-06-21T10:02:11Zcom_10261_135com_10261_4col_10261_388
00925njm 22002777a 4500
dc
Martínez-Cruz, Luis Alfonso
author
Encinar, José Antonio
author
Sevilla, Paz
author
Oyenarte, Iker
author
Gómez García, Inmaculada
author
Aguado Llera, David
author
García Blanco, Francisco
author
Gómez, Javier
author
Neira, José L.
author
2011
Nucleotide-binding cystathionine -synthase (CBS) domains function as regulatory motifs in several proteins distributed through all kingdoms of life. This function has been proposed based on their affinity for adenosyl- derivatives, although the exact binding mechanisms remain largely unknown. The question of how CBS domains exactly work is relevant because in humans, several genetic diseases have been associated with mutations in those motifs. In this work, we describe the adenosyl-ligand (AMP, ATP, NADP and SAM) properties of the wild-type CBS domain protein MJ0729 from Methanocaldococcus jannaschii by using a combination of spectroscopic techniques (fluorescence, FTIR and FRET). The fluorescence results show that binding to AMP and ATP occurs with an apparent dissociation constant of ∼10 M, and interestingly enough, binding induces protein conformational changes, as shown by FTIR. On the other hand, fluorescence spectra (FRET and steady-state) did not change upon addition of NADP and SAM to MJ0729, suggesting that tryptophan and/or tyrosine residues were not involved in the recognition of those ligands; however, there were changes in the secondary structure of the protein upon addition of NADP and SAM, as shown by FTIR (thus, indicating binding to the nucleotide). Taken together, these results suggest that: (i) the adenosyl ligands bind to MJ0729 in different ways, and (ii) there are changes in the protein secondary structure upon binding of the nucleotides. The Author 2010. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals. permissionsoup.com2010 © The Author 2010. Published by Oxford University Press. All rights reserved.
Protein Engineering, Design and Selection 24: 161- 169 (2011)
http://hdl.handle.net/10261/61877
10.1093/protein/gzq073
Nucleotide-induced conformational transitions in the CBS domain protein MJ0729 of Methanocaldococcus jannaschii