File Download
 
Links for fulltext
(May Require Subscription)
 
Supplementary

Article: ACBP4 and ACBP5, novel Arabidopsis acyl-CoA-binding proteins with kelch motifs that bind oleoyl-CoA
  • Basic View
  • Metadata View
  • XML View
TitleACBP4 and ACBP5, novel Arabidopsis acyl-CoA-binding proteins with kelch motifs that bind oleoyl-CoA
 
AuthorsLeung, KC2
Li, HY2 1
Mishra, G2
Chye, ML2
 
Keywords(His) 6-tagged recombinant proteins
ACBP gene family
Acyl-CoA-binding domain
Lipid metabolism
Lipid transfer
Site-directed mutagenesis
 
Issue Date2004
 
PublisherSpringer Verlag Dordrecht. The Journal's web site is located at http://springerlink.metapress.com/openurl.asp?genre=journal&issn=0167-4412
 
CitationPlant Molecular Biology, 2004, v. 55 n. 2, p. 297-309 [How to Cite?]
DOI: http://dx.doi.org/10.1007/s11103-004-0642-z
 
AbstractIn plants, fatty acids synthesized in the chloroplasts are exported as acyl-CoA esters to the endoplasmic reticulum (ER). Cytosolic 10-kDa acyl-CoA-binding proteins (ACBPs), prevalent in eukaryotes, are involved in the storage and intracellular transport of acyl-CoAs. We have previously characterized Arabidopsis thaliana cDNAs encoding membrane-associated ACBPs with ankyrin repeats, designated ACBP1 and ACBP2, which show conservation to cytosolic ACBPs at the acyl-CoA-binding domain. Analysis of the Arabidopsis genome has revealed the presence of three more genes encoding putative proteins with acyl-CoA-binding domains, designated ACBP3, ACBP4 and ACBP5. Homologues of ACBP1 to ACBP5 have not been reported in any other organism. We show by reverse-transcriptase polymerase chain reaction (RT-PCR) analysis that ACBP3, ACBP4 and ACBP5 are expressed in all plant organs, like ACBP1 and ACBP2. ACBP4 and ACBP5 that share 81.4% identity and which contain kelch motifs were further investigated. To demonstrate their function in binding acyl-CoA, we have expressed them as (His) 6-tagged recombinant proteins in Escherichia coli for in vitro binding assays. Both (His) 6-ACBP4 and (His) 6-ACBP5 bind [ 14C]oleoyl-CoA with high affinity, [ 14C]palmitoyl-CoA with lower affinity and did not bind [ 14C]arachidonyl-CoA. Eight mutant forms of each protein with single amino acid substitutions within the acyl-CoA-binding domain were produced and analyzed. On binding assays, all mutants were impaired in oleoyl-CoA binding. Hence, these novel ACBPs with kelch motifs have functional acyl-CoA-binding domains that bind oleoyl-CoA. Their predicted cytosol localization suggests that they could maintain an oleoyl-CoA pool in the cytosol or transport oleoyl-CoA from the plastids to the ER in plant lipid metabolism.
 
ISSN0167-4412
2013 Impact Factor: 4.072
 
DOIhttp://dx.doi.org/10.1007/s11103-004-0642-z
 
ISI Accession Number IDWOS:000225690100011
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorLeung, KC
 
dc.contributor.authorLi, HY
 
dc.contributor.authorMishra, G
 
dc.contributor.authorChye, ML
 
dc.date.accessioned2010-09-06T06:06:45Z
 
dc.date.available2010-09-06T06:06:45Z
 
dc.date.issued2004
 
dc.description.abstractIn plants, fatty acids synthesized in the chloroplasts are exported as acyl-CoA esters to the endoplasmic reticulum (ER). Cytosolic 10-kDa acyl-CoA-binding proteins (ACBPs), prevalent in eukaryotes, are involved in the storage and intracellular transport of acyl-CoAs. We have previously characterized Arabidopsis thaliana cDNAs encoding membrane-associated ACBPs with ankyrin repeats, designated ACBP1 and ACBP2, which show conservation to cytosolic ACBPs at the acyl-CoA-binding domain. Analysis of the Arabidopsis genome has revealed the presence of three more genes encoding putative proteins with acyl-CoA-binding domains, designated ACBP3, ACBP4 and ACBP5. Homologues of ACBP1 to ACBP5 have not been reported in any other organism. We show by reverse-transcriptase polymerase chain reaction (RT-PCR) analysis that ACBP3, ACBP4 and ACBP5 are expressed in all plant organs, like ACBP1 and ACBP2. ACBP4 and ACBP5 that share 81.4% identity and which contain kelch motifs were further investigated. To demonstrate their function in binding acyl-CoA, we have expressed them as (His) 6-tagged recombinant proteins in Escherichia coli for in vitro binding assays. Both (His) 6-ACBP4 and (His) 6-ACBP5 bind [ 14C]oleoyl-CoA with high affinity, [ 14C]palmitoyl-CoA with lower affinity and did not bind [ 14C]arachidonyl-CoA. Eight mutant forms of each protein with single amino acid substitutions within the acyl-CoA-binding domain were produced and analyzed. On binding assays, all mutants were impaired in oleoyl-CoA binding. Hence, these novel ACBPs with kelch motifs have functional acyl-CoA-binding domains that bind oleoyl-CoA. Their predicted cytosol localization suggests that they could maintain an oleoyl-CoA pool in the cytosol or transport oleoyl-CoA from the plastids to the ER in plant lipid metabolism.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationPlant Molecular Biology, 2004, v. 55 n. 2, p. 297-309 [How to Cite?]
DOI: http://dx.doi.org/10.1007/s11103-004-0642-z
 
dc.identifier.doihttp://dx.doi.org/10.1007/s11103-004-0642-z
 
dc.identifier.epage309
 
dc.identifier.hkuros96630
 
dc.identifier.isiWOS:000225690100011
 
dc.identifier.issn0167-4412
2013 Impact Factor: 4.072
 
dc.identifier.issue2
 
dc.identifier.openurl
 
dc.identifier.pmid15604682
 
dc.identifier.scopuseid_2-s2.0-12544254338
 
dc.identifier.spage297
 
dc.identifier.urihttp://hdl.handle.net/10722/68685
 
dc.identifier.volume55
 
dc.languageeng
 
dc.publisherSpringer Verlag Dordrecht. The Journal's web site is located at http://springerlink.metapress.com/openurl.asp?genre=journal&issn=0167-4412
 
dc.publisher.placeNetherlands
 
dc.relation.ispartofPlant Molecular Biology
 
dc.relation.referencesReferences in Scopus
 
dc.subject(His) 6-tagged recombinant proteins
 
dc.subjectACBP gene family
 
dc.subjectAcyl-CoA-binding domain
 
dc.subjectLipid metabolism
 
dc.subjectLipid transfer
 
dc.subjectSite-directed mutagenesis
 
dc.titleACBP4 and ACBP5, novel Arabidopsis acyl-CoA-binding proteins with kelch motifs that bind oleoyl-CoA
 
dc.typeArticle
 
<?xml encoding="utf-8" version="1.0"?>
<item><contributor.author>Leung, KC</contributor.author>
<contributor.author>Li, HY</contributor.author>
<contributor.author>Mishra, G</contributor.author>
<contributor.author>Chye, ML</contributor.author>
<date.accessioned>2010-09-06T06:06:45Z</date.accessioned>
<date.available>2010-09-06T06:06:45Z</date.available>
<date.issued>2004</date.issued>
<identifier.citation>Plant Molecular Biology, 2004, v. 55 n. 2, p. 297-309</identifier.citation>
<identifier.issn>0167-4412</identifier.issn>
<identifier.uri>http://hdl.handle.net/10722/68685</identifier.uri>
<description.abstract>In plants, fatty acids synthesized in the chloroplasts are exported as acyl-CoA esters to the endoplasmic reticulum (ER). Cytosolic 10-kDa acyl-CoA-binding proteins (ACBPs), prevalent in eukaryotes, are involved in the storage and intracellular transport of acyl-CoAs. We have previously characterized Arabidopsis thaliana cDNAs encoding membrane-associated ACBPs with ankyrin repeats, designated ACBP1 and ACBP2, which show conservation to cytosolic ACBPs at the acyl-CoA-binding domain. Analysis of the Arabidopsis genome has revealed the presence of three more genes encoding putative proteins with acyl-CoA-binding domains, designated ACBP3, ACBP4 and ACBP5. Homologues of ACBP1 to ACBP5 have not been reported in any other organism. We show by reverse-transcriptase polymerase chain reaction (RT-PCR) analysis that ACBP3, ACBP4 and ACBP5 are expressed in all plant organs, like ACBP1 and ACBP2. ACBP4 and ACBP5 that share 81.4% identity and which contain kelch motifs were further investigated. To demonstrate their function in binding acyl-CoA, we have expressed them as (His) 6-tagged recombinant proteins in Escherichia coli for in vitro binding assays. Both (His) 6-ACBP4 and (His) 6-ACBP5 bind [ 14C]oleoyl-CoA with high affinity, [ 14C]palmitoyl-CoA with lower affinity and did not bind [ 14C]arachidonyl-CoA. Eight mutant forms of each protein with single amino acid substitutions within the acyl-CoA-binding domain were produced and analyzed. On binding assays, all mutants were impaired in oleoyl-CoA binding. Hence, these novel ACBPs with kelch motifs have functional acyl-CoA-binding domains that bind oleoyl-CoA. Their predicted cytosol localization suggests that they could maintain an oleoyl-CoA pool in the cytosol or transport oleoyl-CoA from the plastids to the ER in plant lipid metabolism.</description.abstract>
<language>eng</language>
<publisher>Springer Verlag Dordrecht. The Journal&apos;s web site is located at http://springerlink.metapress.com/openurl.asp?genre=journal&amp;issn=0167-4412</publisher>
<relation.ispartof>Plant Molecular Biology</relation.ispartof>
<subject>(His) 6-tagged recombinant proteins</subject>
<subject>ACBP gene family</subject>
<subject>Acyl-CoA-binding domain</subject>
<subject>Lipid metabolism</subject>
<subject>Lipid transfer</subject>
<subject>Site-directed mutagenesis</subject>
<title>ACBP4 and ACBP5, novel Arabidopsis acyl-CoA-binding proteins with kelch motifs that bind oleoyl-CoA</title>
<type>Article</type>
<identifier.openurl>http://library.hku.hk:4550/resserv?sid=HKU:IR&amp;issn=0167-4412&amp;volume=55&amp;spage=297&amp;epage=309&amp;date=2004&amp;atitle=ACBP4+and+ACBP5,+novel+Arabidopsis+acyl-CoA+binding+proteins,+with+kelch+motifs+that+bind+oleoyl-CoA</identifier.openurl>
<description.nature>Link_to_subscribed_fulltext</description.nature>
<identifier.doi>10.1007/s11103-004-0642-z</identifier.doi>
<identifier.pmid>15604682</identifier.pmid>
<identifier.scopus>eid_2-s2.0-12544254338</identifier.scopus>
<identifier.hkuros>96630</identifier.hkuros>
<relation.references>http://www.scopus.com/mlt/select.url?eid=2-s2.0-12544254338&amp;selection=ref&amp;src=s&amp;origin=recordpage</relation.references>
<identifier.volume>55</identifier.volume>
<identifier.issue>2</identifier.issue>
<identifier.spage>297</identifier.spage>
<identifier.epage>309</identifier.epage>
<identifier.isi>WOS:000225690100011</identifier.isi>
<publisher.place>Netherlands</publisher.place>
</item>
Author Affiliations
  1. South China Institute of Botany Chinese Academy of Sciences
  2. The University of Hong Kong