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dc.contributor.authorTrujillo, Uldaeliz
dc.contributor.authorVázquez-Rosa, Edwin
dc.contributor.authorOyola-Robles, Delise
dc.contributor.authorStagg, Loren J.
dc.contributor.authorVassallo, David A.
dc.contributor.authorVega, Irving E.
dc.contributor.authorArold, Stefan T.
dc.contributor.authorBaerga-Ortiz, Abel
dc.date.accessioned2014-08-27T09:46:25Z
dc.date.available2014-08-27T09:46:25Z
dc.date.issued2013-02-28
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.
dc.identifier.issn19326203
dc.identifier.pmid23469090
dc.identifier.doi10.1371/journal.pone.0057859
dc.identifier.urihttp://hdl.handle.net/10754/325312
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.
dc.language.isoen
dc.publisherPublic Library of Science (PLoS)
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.
dc.rightsArchived with thanks to PLoS ONE
dc.subjectacyl carrier protein
dc.subjectfatty acid synthase
dc.subjectpolyunsaturated fatty acid synthase
dc.subjectunclassified drug
dc.subjectalgorithm
dc.subjectbiocatalysis
dc.subjectcircular dichroism
dc.subjectcontrolled study
dc.subjectDNA sequence
dc.subjectelectrospray mass spectrometry
dc.subjectgel filtration chromatography
dc.subjectmolecular dynamics
dc.subjectmolecular model
dc.subjectmolecular weight
dc.subjectnucleotide sequence
dc.subjectPhotobacterium phosphoreum
dc.subjectprotein conformation
dc.subjectprotein denaturation
dc.subjectprotein determination
dc.subjectprotein domain
dc.subjectprotein expression
dc.subjectprotein function
dc.subjectprotein quaternary structure
dc.subjectprotein stability
dc.subjectprotein unfolding
dc.subjectsequence analysis
dc.subjectthermal analysis
dc.subjectX ray crystallography
dc.subjectAcyl Carrier Protein
dc.subjectAlgorithms
dc.subjectAmino Acid Sequence
dc.subjectFatty Acid Synthetase Complex
dc.subjectModels, Molecular
dc.subjectMolecular Sequence Data
dc.subjectPeptide Fragments
dc.subjectPhotobacterium
dc.subjectProtein Denaturation
dc.subjectProtein Structure, Tertiary
dc.subjectSolutions
dc.subjectTemperature
dc.titleSolution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentComputational Bioscience Research Center (CBRC)
dc.contributor.departmentStructural Biology and Engineering
dc.identifier.journalPLoS ONE
dc.identifier.pmcidPMC3585217
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionDepartment of Biochemistry, University of Puerto Rico - Medical Sciences Campus, San Juan, Puerto Rico
dc.contributor.institutionDepartment of Biology, University of Puerto Rico - Rio Piedras Campus, San Juan, Puerto Rico
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 States
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)
kaust.personArold, Stefan T.
refterms.dateFOA2018-06-13T14:55:16Z


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