Solution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration

Handle URI:
http://hdl.handle.net/10754/325312
Title:
Solution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration
Authors:
Trujillo, Uldaeliz; Vázquez-Rosa, Edwin; Oyola-Robles, Delise; Stagg, Loren J.; Vassallo, David A.; Vega, Irving E.; Arold, Stefan T. ( 0000-0001-5278-0668 ) ; Baerga-Ortiz, Abel
Abstract:
The polyunsaturated fatty acid (PUFA) synthases from deep-sea bacteria invariably contain multiple acyl carrier protein (ACP) domains in tandem. This conserved tandem arrangement has been implicated in both amplification of fatty acid production (additive effect) and in structural stabilization of the multidomain protein (synergistic effect). While the more accepted model is one in which domains act independently, recent reports suggest that ACP domains may form higher oligomers. Elucidating the three-dimensional structure of tandem arrangements may therefore give important insights into the functional relevance of these structures, and hence guide bioengineering strategies. In an effort to elucidate the three-dimensional structure of tandem repeats from deep-sea anaerobic bacteria, we have expressed and purified a fragment consisting of five tandem ACP domains from the PUFA synthase from Photobacterium profundum. Analysis of the tandem ACP fragment by analytical gel filtration chromatography showed a retention time suggestive of a multimeric protein. However, small angle X-ray scattering (SAXS) revealed that the multi-ACP fragment is an elongated monomer which does not form a globular unit. Stokes radii calculated from atomic monomeric SAXS models were comparable to those measured by analytical gel filtration chromatography, showing that in the gel filtration experiment, the molecular weight was overestimated due to the elongated protein shape. Thermal denaturation monitored by circular dichroism showed that unfolding of the tandem construct was not cooperative, and that the tandem arrangement did not stabilize the protein. Taken together, these data are consistent with an elongated beads-on-a-string arrangement of the tandem ACP domains in PUFA synthases, and speak against synergistic biocatalytic effects promoted by quaternary structuring. Thus, it is possible to envision bioengineering strategies which simply involve the artificial linking of multiple ACP domains for increasing the yield of fatty acids in bacterial cultures. 2013 Trujillo et al.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
Trujillo U, Vázquez-Rosa E, Oyola-Robles D, Stagg LJ, Vassallo DA, et al. (2013) Solution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration. PLoS ONE 8: e57859. doi:10.1371/journal.pone.0057859.
Publisher:
Public Library of Science
Journal:
PLoS ONE
Issue Date:
28-Feb-2013
DOI:
10.1371/journal.pone.0057859
PubMed ID:
23469090
PubMed Central ID:
PMC3585217
Type:
Article
ISSN:
19326203
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorTrujillo, Uldaelizen
dc.contributor.authorVázquez-Rosa, Edwinen
dc.contributor.authorOyola-Robles, Deliseen
dc.contributor.authorStagg, Loren J.en
dc.contributor.authorVassallo, David A.en
dc.contributor.authorVega, Irving E.en
dc.contributor.authorArold, Stefan T.en
dc.contributor.authorBaerga-Ortiz, Abelen
dc.date.accessioned2014-08-27T09:46:25Zen
dc.date.available2014-08-27T09:46:25Zen
dc.date.issued2013-02-28en
dc.identifier.citationTrujillo U, Vázquez-Rosa E, Oyola-Robles D, Stagg LJ, Vassallo DA, et al. (2013) Solution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration. PLoS ONE 8: e57859. doi:10.1371/journal.pone.0057859.en
dc.identifier.issn19326203en
dc.identifier.pmid23469090en
dc.identifier.doi10.1371/journal.pone.0057859en
dc.identifier.urihttp://hdl.handle.net/10754/325312en
dc.description.abstractThe polyunsaturated fatty acid (PUFA) synthases from deep-sea bacteria invariably contain multiple acyl carrier protein (ACP) domains in tandem. This conserved tandem arrangement has been implicated in both amplification of fatty acid production (additive effect) and in structural stabilization of the multidomain protein (synergistic effect). While the more accepted model is one in which domains act independently, recent reports suggest that ACP domains may form higher oligomers. Elucidating the three-dimensional structure of tandem arrangements may therefore give important insights into the functional relevance of these structures, and hence guide bioengineering strategies. In an effort to elucidate the three-dimensional structure of tandem repeats from deep-sea anaerobic bacteria, we have expressed and purified a fragment consisting of five tandem ACP domains from the PUFA synthase from Photobacterium profundum. Analysis of the tandem ACP fragment by analytical gel filtration chromatography showed a retention time suggestive of a multimeric protein. However, small angle X-ray scattering (SAXS) revealed that the multi-ACP fragment is an elongated monomer which does not form a globular unit. Stokes radii calculated from atomic monomeric SAXS models were comparable to those measured by analytical gel filtration chromatography, showing that in the gel filtration experiment, the molecular weight was overestimated due to the elongated protein shape. Thermal denaturation monitored by circular dichroism showed that unfolding of the tandem construct was not cooperative, and that the tandem arrangement did not stabilize the protein. Taken together, these data are consistent with an elongated beads-on-a-string arrangement of the tandem ACP domains in PUFA synthases, and speak against synergistic biocatalytic effects promoted by quaternary structuring. Thus, it is possible to envision bioengineering strategies which simply involve the artificial linking of multiple ACP domains for increasing the yield of fatty acids in bacterial cultures. 2013 Trujillo et al.en
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en
dc.rightsArchived with thanks to PLoS ONEen
dc.subjectacyl carrier proteinen
dc.subjectfatty acid synthaseen
dc.subjectpolyunsaturated fatty acid synthaseen
dc.subjectunclassified drugen
dc.subjectalgorithmen
dc.subjectbiocatalysisen
dc.subjectcircular dichroismen
dc.subjectcontrolled studyen
dc.subjectDNA sequenceen
dc.subjectelectrospray mass spectrometryen
dc.subjectgel filtration chromatographyen
dc.subjectmolecular dynamicsen
dc.subjectmolecular modelen
dc.subjectmolecular weighten
dc.subjectnucleotide sequenceen
dc.subjectPhotobacterium phosphoreumen
dc.subjectprotein conformationen
dc.subjectprotein denaturationen
dc.subjectprotein determinationen
dc.subjectprotein domainen
dc.subjectprotein expressionen
dc.subjectprotein functionen
dc.subjectprotein quaternary structureen
dc.subjectprotein stabilityen
dc.subjectprotein unfoldingen
dc.subjectsequence analysisen
dc.subjectthermal analysisen
dc.subjectX ray crystallographyen
dc.subjectAcyl Carrier Proteinen
dc.subjectAlgorithmsen
dc.subjectAmino Acid Sequenceen
dc.subjectFatty Acid Synthetase Complexen
dc.subjectModels, Molecularen
dc.subjectMolecular Sequence Dataen
dc.subjectPeptide Fragmentsen
dc.subjectPhotobacteriumen
dc.subjectProtein Denaturationen
dc.subjectProtein Structure, Tertiaryen
dc.subjectSolutionsen
dc.subjectTemperatureen
dc.titleSolution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configurationen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalPLoS ONEen
dc.identifier.pmcidPMC3585217en
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.institutionDepartment of Biochemistry, University of Puerto Rico - Medical Sciences Campus, San Juan, Puerto Ricoen
dc.contributor.institutionDepartment of Biology, University of Puerto Rico - Rio Piedras Campus, San Juan, Puerto Ricoen
dc.contributor.institutionDepartment of Biochemistry and Molecular Biology and Center for Biomolecular Structure and Function, The University of Texas MD Anderson Cancer Center, Houston, TX, United Statesen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorArold, Stefan T.en

Related articles on PubMed

All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.