Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE

NovP is an S-adenosyl-l-methionine-dependent O-methyltransferase from Streptomyces spheroides (subunit MW = 29 967 Da). Recombinant N-terminally His-tagged NovP crystallizes in space group P2, with approximate unit-cell parameters a = 51.81, b = 46.04, c = 61.22 Å, β = 105.0°, giving a solvent conte...

ver descrição completa

Detalhes bibliográficos
Autores: Usón, Isabel, Stevenson, Clare E. M., Lawson, David M., Sheldrick, George M.
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2007
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/113906
Acesso em linha:http://hdl.handle.net/10261/113906
Access Level:acceso abierto
Palavra-chave:Antibiotic biosynthesis
Free lunch algorithm
SHELXE
NovP
O-methyltransferase
Chain tracing
id ES_a28bd2b30d8ca2aa1b7ed72afdd706a0
oai_identifier_str oai:digital.csic.es:10261/113906
network_acronym_str ES
network_name_str España
repository_id_str
spelling Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXEUsón, IsabelStevenson, Clare E. M.Lawson, David M.Sheldrick, George M.Antibiotic biosynthesisFree lunch algorithmSHELXENovPO-methyltransferaseChain tracingNovP is an S-adenosyl-l-methionine-dependent O-methyltransferase from Streptomyces spheroides (subunit MW = 29 967 Da). Recombinant N-terminally His-tagged NovP crystallizes in space group P2, with approximate unit-cell parameters a = 51.81, b = 46.04, c = 61.22 Å, β = 105.0°, giving a solvent content of 44% for a single copy of the His-tagged protomer per asymmetric unit. Native synchrotron data to a resolution of 1.35 Å were combined with three other native data sets collected at lower resolution (both in-house and at the synchrotron) for the sake of completeness and better scaling. Data to 2.45 Å resolution were subsequently recorded in-house from a single mercury derivative. Three partial mercury sites could be located with SHELXD, but the resulting phases had a mean error of about 81° and in our hands did not yield an interpretable map using standard automated software. Nevertheless, the structure of NovP could be solved by first tracing a small part of the structure by hand and then extrapolating within and beyond the experimental resolution limit using the 'free lunch algorithm' in SHELXE. The resulting phases have a mean phase error of 17° relative to a refined model. © International Union of Crystallography 2007.IU and GMS are grateful to the European Union Integrated Project BIOXHIT for support. IU thanks the Spanish MEC for financial support (grant BIO2003-06653). CEMS and DML would like to acknowledge the BBSRC for financial support through responsive mode funding (ref. B19400) and the Core Strategic Grant to the John Innes Centre. GMS thanks the Fonds der Chemischen Industrie for supportPeer ReviewedBlackwell Publishing2015201520072015info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/113906reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1107/S0907444907042230info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1139062026-05-22T06:33:51Z
dc.title.none.fl_str_mv Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
title Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
spellingShingle Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
Usón, Isabel
Antibiotic biosynthesis
Free lunch algorithm
SHELXE
NovP
O-methyltransferase
Chain tracing
title_short Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
title_full Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
title_fullStr Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
title_full_unstemmed Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
title_sort Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
dc.creator.none.fl_str_mv Usón, Isabel
Stevenson, Clare E. M.
Lawson, David M.
Sheldrick, George M.
author Usón, Isabel
author_facet Usón, Isabel
Stevenson, Clare E. M.
Lawson, David M.
Sheldrick, George M.
author_role author
author2 Stevenson, Clare E. M.
Lawson, David M.
Sheldrick, George M.
author2_role author
author
author
dc.subject.none.fl_str_mv Antibiotic biosynthesis
Free lunch algorithm
SHELXE
NovP
O-methyltransferase
Chain tracing
topic Antibiotic biosynthesis
Free lunch algorithm
SHELXE
NovP
O-methyltransferase
Chain tracing
description NovP is an S-adenosyl-l-methionine-dependent O-methyltransferase from Streptomyces spheroides (subunit MW = 29 967 Da). Recombinant N-terminally His-tagged NovP crystallizes in space group P2, with approximate unit-cell parameters a = 51.81, b = 46.04, c = 61.22 Å, β = 105.0°, giving a solvent content of 44% for a single copy of the His-tagged protomer per asymmetric unit. Native synchrotron data to a resolution of 1.35 Å were combined with three other native data sets collected at lower resolution (both in-house and at the synchrotron) for the sake of completeness and better scaling. Data to 2.45 Å resolution were subsequently recorded in-house from a single mercury derivative. Three partial mercury sites could be located with SHELXD, but the resulting phases had a mean error of about 81° and in our hands did not yield an interpretable map using standard automated software. Nevertheless, the structure of NovP could be solved by first tracing a small part of the structure by hand and then extrapolating within and beyond the experimental resolution limit using the 'free lunch algorithm' in SHELXE. The resulting phases have a mean phase error of 17° relative to a refined model. © International Union of Crystallography 2007.
publishDate 2007
dc.date.none.fl_str_mv 2007
2015
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/113906
url http://hdl.handle.net/10261/113906
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1107/S0907444907042230
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Blackwell Publishing
publisher.none.fl_str_mv Blackwell Publishing
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869415289350782976
score 15,812429

Registos relacionados