IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ A conserved domain of yeast RNA triphosphatase flanking the catalytic core regulates self-association and interaction with the guanylyltransferase component of the mRNA capping apparatus. A conserved domain of yeast RNA triphosphatase flanking the catalytic core regulates self-association and interaction with the guanylyltransferase component of the mRNA capping apparatus. J. Biol. Chem. (0021-9258) 1999 imex curation Lehman K., Schwer B., Ho CK., Rouzankina I., Shuman S. Update to ATPase assay when term added to CV Triphosphatase activity was determined by assaying manganese-dependent 32P-labelled ATP hydrolysis. s-shuman@ski.mskcc.org 23-MAY-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Lehman K., Schwer B., Ho CK., Rouzankina I., Shuman S. Update to ATPase assay when term added to CV Triphosphatase activity was determined by assaying manganese-dependent 32P-labelled ATP hydrolysis. s-shuman@ski.mskcc.org Accepted 2007-JUN-11 by JYOTI. 23-MAY-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1999 Only protein-protein interactions imex curation A conserved domain of yeast RNA triphosphatase flanking the catalytic core regulates self-association and interaction with the guanylyltransferase component of the mRNA capping apparatus. A conserved domain of yeast RNA triphosphatase flanking the catalytic core regulates self-association and interaction with the guanylyltransferase component of the mRNA capping apparatus. J. Biol. Chem. (0021-9258) 1999 imex curation Lehman K., Schwer B., Ho CK., Rouzankina I., Shuman S. Update to ATPase assay when term added to CV Triphosphatase activity was determined by assaying manganese-dependent 32P-labelled ATP hydrolysis. s-shuman@ski.mskcc.org 23-MAY-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation predetermined predetermined participant predetermined Lehman K., Schwer B., Ho CK., Rouzankina I., Shuman S. 15-30% glycerol gradient s-shuman@ski.mskcc.org Accepted 2007-JUN-10 by JYOTI. 23-MAY-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1999 Only protein-protein interactions imex curation cet1_yeast mRNA-capping enzyme subunit beta CET1 mRNA 5'-phosphatase mRNA 5'-triphosphate monophosphatase P1433 YPL228W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSYTDNPPQTKRALSLDDLVNHDENEKVKLQKLSEAANGSRPFAENLESDINQTETGQAAPIDNYKESTGHGSHSQKPKSRKSSNDDEETDTDDEMGASGEINFDSEMDFDYDKQHRNLLSNGSPPMNDGSDANAKLEKPSDDSIHQNSKSDEEQRIPKQGNEGNIASNYITQVPLQKQKQTEKKIAGNAVGSVVKKEEEANAAVDNIFEEKATLQSKKNNIKRDLEVLNEISASSKPSKYRNVPIWAQKWKPTIKALQSINVKDLKIDPSFLNIIPDDDLTKSVQDWVYATIYSIAPELRSFIELEMKFGVIIDAKGPDRVNPPVSSQCVFTELDAHLTPNIDASLFKELSKYIRGISEVTENTGKFSIIESQTRDSVYRVGLSTQRPRFLRMSTDIKTGRVGQFIEKRHVAQLLLYSPKDSYDVKISLNLELPVPDNDPPEKYKSQSPISERTKDRVSYIHNDSCTRIDITKVENHNQNSKSRQSETTHEVELEINTPALLNAFDNITNDSKEYASLIRTFLNNGTIIRRKLSSLSYEIFEGSKKVM 2B946CC53D66F824 2B946CC53D66F824 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mce1_yeast mRNA-capping enzyme subunit alpha CEG1 YGL130W G2853 mRNA guanylyltransferase GTP--RNA guanylyltransferase protein protein yeast Saccharomyces cerevisiae Baker's yeast MVLAMESRVAPEIPGLIQPGNVTQDLKMMVCKLLNSPKPTKTFPGSQPVSFQHSDVEEKLLAHDYYVCEKTDGLRVLMFIVINPVTGEQGCFMIDRENNYYLVNGFRFPRLPQKKKEELLETLQDGTLLDGELVIQTNPMTKLQELRYLMFDCLAINGRCLTQSPTSSRLAHLGKEFFKPYFDLRAAYPNRCTTFPFKISMKHMDFSYQLVKVAKSLDKLPHLSDGLIFTPVKAPYTAGGKDSLLLKWKPEQENTVDFKLILDIPMVEDPSLPKDDRNRWYYNYDVKPVFSLYVWQGGADVNSRLKHFDQPFDRKEFEILERTYRKFAELSVSDEEWQNLKNLEQPLNGRIVECAKNQETGAWEMLRFRDDKLNGNHTSVVQKVLESINDSVSLEDLEEIVGDIKRCWDERRANMAGGSGRPLPSQSQNATLSTSKPVHSQPPSNDKEPKYVDEDDWSD F7F5A3E59EEFA15F F7F5A3E59EEFA15F cet1-1 1 3 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphotransfer phosphotransfer reaction phosphotransfer The instructive finding was that Cet1(276-549)p also retained full triphosphatase function in vitro (Fig. 4B). 4B cet1-2 2 3 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction The Delta 200 triphosphatase (a 43-kDa polypeptide) sedimented as a discrete peak (fraction 15) just slightly heavier than the BSA peak (68-kDa). 7A cet1-ceg1-1 2 5 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 3 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association The sedimentation coefficient of the Cet1(201-549)p-Ceg1p complex was 7.3. This value suggested a heterotrimeric subunit structure which, given that Cet1(201-549)p per se sediments as a dimer, whereas Ceg1p alone is monomeric (see below), likely consists of two molecules of Cet1(201-549)p and one molecule of Ceg1p. 7B, 9 The sedimentation coefficient of the Cet1(230-549)p-Ceg1p complex was 7.1. Further truncation of Cet1 to 245-549 resulted in the formation of a multimer, although some protein did still form the heterotrimer with Ceg1.