2024-03-28T09:58:51Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/104882020-04-07T09:36:23Zcom_10261_33com_10261_5col_10261_286
C-Terminal Truncation of α 1,6-Fucosyltransferase from Rhizobium Sp. does not Annul the Transferase Activity of the Enzyme
Bastida, Agatha
Fernández-Mayoralas, Alfonso
García-Junceda, Eduardo
1,6-Fucosyltransferase
Rhizobium Sp.
Activity of the enzyme
C-terminal
protein
TMpred
6 pages.-- PMID: 11814863 [PubMed].
Recently we have over-expressed the enzyme a 1,6-fucosyltransferase from Rhizobium sp. in Escherichia coli. In this heterologous system the enzyme was mainly expressed as inclusion bodies and the one that was expressed soluble showed a shortlasting activity in solution due to precipitation of the protein. A structural analysis of the sequence using the TMpred program
predicted a highly hydrophobic region of 19 aa close to the C-terminal of the protein. In order to investigate the influence of this region on the formation of inclusion bodies and the precipitation from solution, we cloned a truncated version of the protein where a C-terminal fragment of 65 aa, including the predicted transmembrane-like region, was removed. The resulting protein was expressed in a soluble form without formation of inclusion bodies. The truncated protein catalyzed the transfer of a fucopyranosyl moiety from GDP-b-l-Fucose to chitobiose. Comparison of the acceptor specificity between the truncated a 1,6-fucosyltransferase and the wild-type enzyme, showed a similar behavior for both enzymes. Our results indicate that the active center is not located in the C-terminal extreme of the protein in contrast to the case of the mammalian glycosyltransferases. Also, these results indicate that the a-6-motif III is not directly involved in the catalytic activity of the enzyme.
This work was supported by the Spanish DGES (Grant PB96-0828) and Comunidad de Madrid (Grant 07B/0027/1999).
Peer reviewed
2009-02-10T09:37:13Z
2009-02-10T09:37:13Z
2002-03
artículo
http://purl.org/coar/resource_type/c_6501
Bioorganic & Medicinal Chemistry 12(8): 1817-1834 (2002)
0968-0896
http://hdl.handle.net/10261/10488
10.1016/S0968-0896(01)00327-3
en
http://dx.doi.org/10.1016/S0968-0896(01)00327-3
open
458854 bytes
application/pdf
Pergamon Press