Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis.
Identifieur interne : 002705 ( PubMed/Checkpoint ); précédent : 002704; suivant : 002706Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis.
Auteurs : K M Parkhurst ; R E Hileman ; D. Saha ; N K Gupta ; L J ParkhurstSource :
- Biochemistry [ 0006-2960 ] ; 1994.
Descripteurs français
- KwdFr :
- ARN de transfert de la méthionine (métabolisme), ARN messager (métabolisme), Animaux, Biosynthèse des protéines, Codon, Colorants fluorescents, Données de séquences moléculaires, Facteur-2 d'initiation eucaryote (métabolisme), Facteurs d'échange de nucléotides guanyliques, Guanosine triphosphate (métabolisme), Polarisation de fluorescence, Protéines (métabolisme), Relation structure-activité, Ribosomes (métabolisme), Sites de fixation, Séquence nucléotidique, Thermodynamique.
- MESH :
- métabolisme : ARN de transfert de la méthionine, ARN messager, Facteur-2 d'initiation eucaryote, Guanosine triphosphate, Protéines, Ribosomes.
- Animaux, Biosynthèse des protéines, Codon, Colorants fluorescents, Données de séquences moléculaires, Facteurs d'échange de nucléotides guanyliques, Polarisation de fluorescence, Relation structure-activité, Sites de fixation, Séquence nucléotidique, Thermodynamique.
English descriptors
- KwdEn :
- Animals, Base Sequence, Binding Sites, Codon, Eukaryotic Initiation Factor-2 (metabolism), Fluorescence Polarization, Fluorescent Dyes, Guanine Nucleotide Exchange Factors, Guanosine Triphosphate (metabolism), Molecular Sequence Data, Protein Biosynthesis, Proteins (metabolism), RNA, Messenger (metabolism), RNA, Transfer, Met (metabolism), Ribosomes (metabolism), Structure-Activity Relationship, Thermodynamics.
- MESH :
- chemical , metabolism : Eukaryotic Initiation Factor-2, Guanosine Triphosphate, Proteins, RNA, Messenger, RNA, Transfer, Met.
- chemical : Codon, Fluorescent Dyes, Guanine Nucleotide Exchange Factors.
- metabolism : Ribosomes.
- Animals, Base Sequence, Binding Sites, Fluorescence Polarization, Molecular Sequence Data, Protein Biosynthesis, Structure-Activity Relationship, Thermodynamics.
Abstract
The first step in mammalian protein synthesis is the formation of the 40S initiation complex, composed of the 40S ribosomal subunit (R), mRNA (M, here, a 10-mer oligoribonucleotide analogue containing the initiation codon), and the quaternary complex (Q, composed of eIF-2, GTP, Met-tRNA(fMet), and the ancillary protein factor Co-eIF-2C). The interdependence of the binding of R, M, and Q in forming the 40S complex is currently unclear. We have determined the thermodynamic parameters that characterize these interactions. The binary constants for R+M and Q+M were determined spectroscopically, measuring changes in the anisotropy of the fluorescence emission of 3'-fluorescein labeled M. The other binary constant, for Q+R, and the ternary constant were determined from Millipore filtration assays using radiolabeled Met-tRNA(fMet). The association constants for the binary reactions were as follows: Ka(Q,M) < or = 0.14 x 10(6) M-1, Ka(R,M) = 1.78 x 10(6) M-1, and Ka(Q,R) = 0.94 x 10(6) M-1. The binding of Q to R.M was markedly greater than that of Q to R [Ka(Q,R.M)/Ka(Q,R) > 62]. High cooperativity for this interaction occurs in either a single-site model or in lattice models for the binding of M to R. Data obtained using five other RNA 10-mers, each with the sequence altered at the AUG codon, suggest that this cooperativity is AUG dependent. The data are consistent with a scheme in which mRNA and Q bind independently to the 40S ribosome, but when the AUG codon is properly aligned with Q, a conformational change results in a 2.4 kcal/mol stabilization of the complex.
DOI: 10.1021/bi00254a028
PubMed: 7999777
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
pubmed:7999777Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis.</title>
<author><name sortKey="Parkhurst, K M" sort="Parkhurst, K M" uniqKey="Parkhurst K" first="K M" last="Parkhurst">K M Parkhurst</name>
<affiliation><nlm:affiliation>Department of Chemistry, University of Nebraska--Lincoln, 68588-0304.</nlm:affiliation>
<wicri:noCountry code="subField">68588-0304</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Hileman, R E" sort="Hileman, R E" uniqKey="Hileman R" first="R E" last="Hileman">R E Hileman</name>
</author>
<author><name sortKey="Saha, D" sort="Saha, D" uniqKey="Saha D" first="D" last="Saha">D. Saha</name>
</author>
<author><name sortKey="Gupta, N K" sort="Gupta, N K" uniqKey="Gupta N" first="N K" last="Gupta">N K Gupta</name>
</author>
<author><name sortKey="Parkhurst, L J" sort="Parkhurst, L J" uniqKey="Parkhurst L" first="L J" last="Parkhurst">L J Parkhurst</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="1994">1994</date>
<idno type="RBID">pubmed:7999777</idno>
<idno type="pmid">7999777</idno>
<idno type="doi">10.1021/bi00254a028</idno>
<idno type="wicri:Area/PubMed/Corpus">002848</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002848</idno>
<idno type="wicri:Area/PubMed/Curation">002848</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002848</idno>
<idno type="wicri:Area/PubMed/Checkpoint">002705</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002705</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis.</title>
<author><name sortKey="Parkhurst, K M" sort="Parkhurst, K M" uniqKey="Parkhurst K" first="K M" last="Parkhurst">K M Parkhurst</name>
<affiliation><nlm:affiliation>Department of Chemistry, University of Nebraska--Lincoln, 68588-0304.</nlm:affiliation>
<wicri:noCountry code="subField">68588-0304</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Hileman, R E" sort="Hileman, R E" uniqKey="Hileman R" first="R E" last="Hileman">R E Hileman</name>
</author>
<author><name sortKey="Saha, D" sort="Saha, D" uniqKey="Saha D" first="D" last="Saha">D. Saha</name>
</author>
<author><name sortKey="Gupta, N K" sort="Gupta, N K" uniqKey="Gupta N" first="N K" last="Gupta">N K Gupta</name>
</author>
<author><name sortKey="Parkhurst, L J" sort="Parkhurst, L J" uniqKey="Parkhurst L" first="L J" last="Parkhurst">L J Parkhurst</name>
</author>
</analytic>
<series><title level="j">Biochemistry</title>
<idno type="ISSN">0006-2960</idno>
<imprint><date when="1994" type="published">1994</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Binding Sites</term>
<term>Codon</term>
<term>Eukaryotic Initiation Factor-2 (metabolism)</term>
<term>Fluorescence Polarization</term>
<term>Fluorescent Dyes</term>
<term>Guanine Nucleotide Exchange Factors</term>
<term>Guanosine Triphosphate (metabolism)</term>
<term>Molecular Sequence Data</term>
<term>Protein Biosynthesis</term>
<term>Proteins (metabolism)</term>
<term>RNA, Messenger (metabolism)</term>
<term>RNA, Transfer, Met (metabolism)</term>
<term>Ribosomes (metabolism)</term>
<term>Structure-Activity Relationship</term>
<term>Thermodynamics</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN de transfert de la méthionine (métabolisme)</term>
<term>ARN messager (métabolisme)</term>
<term>Animaux</term>
<term>Biosynthèse des protéines</term>
<term>Codon</term>
<term>Colorants fluorescents</term>
<term>Données de séquences moléculaires</term>
<term>Facteur-2 d'initiation eucaryote (métabolisme)</term>
<term>Facteurs d'échange de nucléotides guanyliques</term>
<term>Guanosine triphosphate (métabolisme)</term>
<term>Polarisation de fluorescence</term>
<term>Protéines (métabolisme)</term>
<term>Relation structure-activité</term>
<term>Ribosomes (métabolisme)</term>
<term>Sites de fixation</term>
<term>Séquence nucléotidique</term>
<term>Thermodynamique</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Eukaryotic Initiation Factor-2</term>
<term>Guanosine Triphosphate</term>
<term>Proteins</term>
<term>RNA, Messenger</term>
<term>RNA, Transfer, Met</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>Codon</term>
<term>Fluorescent Dyes</term>
<term>Guanine Nucleotide Exchange Factors</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Ribosomes</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN de transfert de la méthionine</term>
<term>ARN messager</term>
<term>Facteur-2 d'initiation eucaryote</term>
<term>Guanosine triphosphate</term>
<term>Protéines</term>
<term>Ribosomes</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Binding Sites</term>
<term>Fluorescence Polarization</term>
<term>Molecular Sequence Data</term>
<term>Protein Biosynthesis</term>
<term>Structure-Activity Relationship</term>
<term>Thermodynamics</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Biosynthèse des protéines</term>
<term>Codon</term>
<term>Colorants fluorescents</term>
<term>Données de séquences moléculaires</term>
<term>Facteurs d'échange de nucléotides guanyliques</term>
<term>Polarisation de fluorescence</term>
<term>Relation structure-activité</term>
<term>Sites de fixation</term>
<term>Séquence nucléotidique</term>
<term>Thermodynamique</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">The first step in mammalian protein synthesis is the formation of the 40S initiation complex, composed of the 40S ribosomal subunit (R), mRNA (M, here, a 10-mer oligoribonucleotide analogue containing the initiation codon), and the quaternary complex (Q, composed of eIF-2, GTP, Met-tRNA(fMet), and the ancillary protein factor Co-eIF-2C). The interdependence of the binding of R, M, and Q in forming the 40S complex is currently unclear. We have determined the thermodynamic parameters that characterize these interactions. The binary constants for R+M and Q+M were determined spectroscopically, measuring changes in the anisotropy of the fluorescence emission of 3'-fluorescein labeled M. The other binary constant, for Q+R, and the ternary constant were determined from Millipore filtration assays using radiolabeled Met-tRNA(fMet). The association constants for the binary reactions were as follows: Ka(Q,M) < or = 0.14 x 10(6) M-1, Ka(R,M) = 1.78 x 10(6) M-1, and Ka(Q,R) = 0.94 x 10(6) M-1. The binding of Q to R.M was markedly greater than that of Q to R [Ka(Q,R.M)/Ka(Q,R) > 62]. High cooperativity for this interaction occurs in either a single-site model or in lattice models for the binding of M to R. Data obtained using five other RNA 10-mers, each with the sequence altered at the AUG codon, suggest that this cooperativity is AUG dependent. The data are consistent with a scheme in which mRNA and Q bind independently to the 40S ribosome, but when the AUG codon is properly aligned with Q, a conformational change results in a 2.4 kcal/mol stabilization of the complex.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">7999777</PMID>
<DateCompleted><Year>1995</Year>
<Month>01</Month>
<Day>24</Day>
</DateCompleted>
<DateRevised><Year>2019</Year>
<Month>06</Month>
<Day>13</Day>
</DateRevised>
<Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-2960</ISSN>
<JournalIssue CitedMedium="Print"><Volume>33</Volume>
<Issue>50</Issue>
<PubDate><Year>1994</Year>
<Month>Dec</Month>
<Day>20</Day>
</PubDate>
</JournalIssue>
<Title>Biochemistry</Title>
<ISOAbbreviation>Biochemistry</ISOAbbreviation>
</Journal>
<ArticleTitle>Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis.</ArticleTitle>
<Pagination><MedlinePgn>15168-77</MedlinePgn>
</Pagination>
<Abstract><AbstractText>The first step in mammalian protein synthesis is the formation of the 40S initiation complex, composed of the 40S ribosomal subunit (R), mRNA (M, here, a 10-mer oligoribonucleotide analogue containing the initiation codon), and the quaternary complex (Q, composed of eIF-2, GTP, Met-tRNA(fMet), and the ancillary protein factor Co-eIF-2C). The interdependence of the binding of R, M, and Q in forming the 40S complex is currently unclear. We have determined the thermodynamic parameters that characterize these interactions. The binary constants for R+M and Q+M were determined spectroscopically, measuring changes in the anisotropy of the fluorescence emission of 3'-fluorescein labeled M. The other binary constant, for Q+R, and the ternary constant were determined from Millipore filtration assays using radiolabeled Met-tRNA(fMet). The association constants for the binary reactions were as follows: Ka(Q,M) < or = 0.14 x 10(6) M-1, Ka(R,M) = 1.78 x 10(6) M-1, and Ka(Q,R) = 0.94 x 10(6) M-1. The binding of Q to R.M was markedly greater than that of Q to R [Ka(Q,R.M)/Ka(Q,R) > 62]. High cooperativity for this interaction occurs in either a single-site model or in lattice models for the binding of M to R. Data obtained using five other RNA 10-mers, each with the sequence altered at the AUG codon, suggest that this cooperativity is AUG dependent. The data are consistent with a scheme in which mRNA and Q bind independently to the 40S ribosome, but when the AUG codon is properly aligned with Q, a conformational change results in a 2.4 kcal/mol stabilization of the complex.</AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Parkhurst</LastName>
<ForeName>K M</ForeName>
<Initials>KM</Initials>
<AffiliationInfo><Affiliation>Department of Chemistry, University of Nebraska--Lincoln, 68588-0304.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Hileman</LastName>
<ForeName>R E</ForeName>
<Initials>RE</Initials>
</Author>
<Author ValidYN="Y"><LastName>Saha</LastName>
<ForeName>D</ForeName>
<Initials>D</Initials>
</Author>
<Author ValidYN="Y"><LastName>Gupta</LastName>
<ForeName>N K</ForeName>
<Initials>NK</Initials>
</Author>
<Author ValidYN="Y"><LastName>Parkhurst</LastName>
<ForeName>L J</ForeName>
<Initials>LJ</Initials>
</Author>
</AuthorList>
<Language>eng</Language>
<GrantList CompleteYN="Y"><Grant><GrantID>DK 36288</GrantID>
<Acronym>DK</Acronym>
<Agency>NIDDK NIH HHS</Agency>
<Country>United States</Country>
</Grant>
<Grant><GrantID>GM 22079</GrantID>
<Acronym>GM</Acronym>
<Agency>NIGMS NIH HHS</Agency>
<Country>United States</Country>
</Grant>
</GrantList>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
<PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType>
</PublicationTypeList>
</Article>
<MedlineJournalInfo><Country>United States</Country>
<MedlineTA>Biochemistry</MedlineTA>
<NlmUniqueID>0370623</NlmUniqueID>
<ISSNLinking>0006-2960</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D003062">Codon</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D015852">Eukaryotic Initiation Factor-2</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D005456">Fluorescent Dyes</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D020662">Guanine Nucleotide Exchange Factors</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D011506">Proteins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D012333">RNA, Messenger</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D012358">RNA, Transfer, Met</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>86-01-1</RegistryNumber>
<NameOfSubstance UI="D006160">Guanosine Triphosphate</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D001665" MajorTopicYN="N">Binding Sites</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D003062" MajorTopicYN="N">Codon</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D015852" MajorTopicYN="N">Eukaryotic Initiation Factor-2</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005454" MajorTopicYN="N">Fluorescence Polarization</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005456" MajorTopicYN="N">Fluorescent Dyes</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D020662" MajorTopicYN="N">Guanine Nucleotide Exchange Factors</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D006160" MajorTopicYN="N">Guanosine Triphosphate</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D014176" MajorTopicYN="Y">Protein Biosynthesis</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D011506" MajorTopicYN="N">Proteins</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012333" MajorTopicYN="N">RNA, Messenger</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012358" MajorTopicYN="N">RNA, Transfer, Met</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012270" MajorTopicYN="N">Ribosomes</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013329" MajorTopicYN="N">Structure-Activity Relationship</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013816" MajorTopicYN="N">Thermodynamics</DescriptorName>
</MeshHeading>
</MeshHeadingList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1994</Year>
<Month>12</Month>
<Day>20</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="medline"><Year>1994</Year>
<Month>12</Month>
<Day>20</Day>
<Hour>0</Hour>
<Minute>1</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>1994</Year>
<Month>12</Month>
<Day>20</Day>
<Hour>0</Hour>
<Minute>0</Minute>
</PubMedPubDate>
</History>
<PublicationStatus>ppublish</PublicationStatus>
<ArticleIdList><ArticleId IdType="pubmed">7999777</ArticleId>
<ArticleId IdType="doi">10.1021/bi00254a028</ArticleId>
</ArticleIdList>
</PubmedData>
</pubmed>
<affiliations><list></list>
<tree><noCountry><name sortKey="Gupta, N K" sort="Gupta, N K" uniqKey="Gupta N" first="N K" last="Gupta">N K Gupta</name>
<name sortKey="Hileman, R E" sort="Hileman, R E" uniqKey="Hileman R" first="R E" last="Hileman">R E Hileman</name>
<name sortKey="Parkhurst, K M" sort="Parkhurst, K M" uniqKey="Parkhurst K" first="K M" last="Parkhurst">K M Parkhurst</name>
<name sortKey="Parkhurst, L J" sort="Parkhurst, L J" uniqKey="Parkhurst L" first="L J" last="Parkhurst">L J Parkhurst</name>
<name sortKey="Saha, D" sort="Saha, D" uniqKey="Saha D" first="D" last="Saha">D. Saha</name>
</noCountry>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/PubMed/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002705 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/PubMed/Checkpoint/biblio.hfd -nk 002705 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= PubMed |étape= Checkpoint |type= RBID |clé= pubmed:7999777 |texte= Thermodynamic characterization of the cooperativity of 40S complex formation during the initiation of eukaryotic protein synthesis. }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/PubMed/Checkpoint/RBID.i -Sk "pubmed:7999777" \ | HfdSelect -Kh $EXPLOR_AREA/Data/PubMed/Checkpoint/biblio.hfd \ | NlmPubMed2Wicri -a MersV1
This area was generated with Dilib version V0.6.33. |