UniProt UniProt http://www.uniprot.org iakhiaeva-2006-1 in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation predetermined predetermined participant predetermined Iakhiaeva E., Bhuiyan SH., Yin J., Zwieb C. zwieb@uthct.edu Protein science : a publication of the Protein Society 2006 Only protein-protein interactions imex curation imex curation iakhiaeva-2006-2 ecoli Escherichia coli (strain K12) density sedimentation cosedimentation through density gradient density sedimentation predetermined predetermined participant predetermined Iakhiaeva E., Bhuiyan SH., Yin J., Zwieb C. A dual-cassette vector that contained the coding regions of the two polypeptides separated by a TEV protease cleavage site and preceded by the gene for thioredoxin.was expressed. Upon cleavage with TEV protease one of the two polypeptides remained as a thioredoxin-fused larger protein..Following digestion with TEV protease fragments were analysed by sucrose gradient centrifugation followed by SDS-PAGE. zwieb@uthct.edu Protein science : a publication of the Protein Society 2006 Only protein-protein interactions imex curation imex curation iakhiaeva-2006-3 in vitro In vitro pull down pull down weight by staining molecular weight estimation by staining weight by staining Iakhiaeva E., Bhuiyan SH., Yin J., Zwieb C. A dual-cassette vector that contained the coding regions of the two polypeptides separated by a TEV protease cleavage site and preceded by the gene for thioredoxin.was expressed. Upon cleavage with TEV protease one of the two polypeptides remained as a thioredoxin-fused larger protein..Following dDigestion with TEV protease fragments were analysed by sucrose gradient centrifugation followed by SDS-PAGE. zwieb@uthct.edu Protein science : a publication of the Protein Society 2006 Only protein-protein interactions imex curation imex curation iakhiaeva-2006-4 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Iakhiaeva E., Bhuiyan SH., Yin J., Zwieb C. A dual-cassette vector that contained the coding regions of the two polypeptides separated by a TEV protease cleavage site and preceded by the gene for thioredoxin.was expressed. Upon cleavage with TEV protease one of the two polypeptides remained as a thioredoxin-fused larger protein..Following dDigestion with TEV protease fragments were analysed by sucrose gradient centrifugation followed by SDS-PAGE. zwieb@uthct.edu Protein science : a publication of the Protein Society 2006 Only protein-protein interactions imex curation imex curation srp68_human Signal recognition particle subunit SRP68 SRP68 Signal recognition particle 68 kDa protein protein protein human Homo sapiens Human MAAEKQVPGGGGGGGSGGGGGSGGGGSGGGRGAGGEENKENERPSAGSKANKEFGDSLSLEILQIIKESQQQHGLRHGDFQRYRGYCSRRQRRLRKTLNFKMGNRHKFTGKKVTEELLTDNRYLLLVLMDAERAWSYAMQLKQEANTEPRKRFHLLSRLRKAVKHAEELERLCESNRVDAKTKLEAQAYTAYLSGMLRFEHQEWKAAIEAFNKCKTIYEKLASAFTEEQAVLYNQRVEEISPNIRYCAYNIGDQSAINELMQMRLRSGGTEGLLAEKLEALITQTRAKQAATMSEVEWRGRTVPVKIDKVRIFLLGLADNEAAIVQAESEETKERLFESMLSECRDAIQVVREELKPDQKQRDYILEGEPGKVSNLQYLHSYLTYIKLSTAIKRNENMAKGLQRALLQQQPEDDSKRSPRPQDLIRLYDIILQNLVELLQLPGLEEDKAFQKEIGLKTLVFKAYRCFFIAQSYVLVKKWSEALVLYDRVLKYANEVNSDAGAFKNSLKDLPDVQELITQVRSEKCSLQAAAILDANDAHQTETSSSQVKDNKPLVERFETFCLDPSLVTKQANLVHFPPGFQPIPCKPLFFDLALNHVAFPPLEDKLEQKTKSGLTGYIKGIFGFRS srp72_human Signal recognition particle subunit SRP72 SRP72 Signal recognition particle 72 kDa protein protein protein human Homo sapiens Human MASGGSGGVSVPALWSEVNRYGQNGDFTRALKTVNKILQINKDDVTALHCKVVCLIQNGSFKEALNVINTHTKVLANNSLSFEKAYCEYRLNRIENALKTIESANQQTDKLKELYGQVLYRLERYDECLAVYRDLVRNSQDDYDEERKTNLSAVVAAQSNWEKVVPENLGLQEGTHELCYNTACALIGQGQLNQAMKILQKAEDLCRRSLSEDTDGTEEDPQAELAIIHGQMAYILQLQGRTEEALQLYNQIIKLKPTDVGLLAVIANNIITINKDQNVFDSKKKVKLTNAEGVEFKLSKKQLQAIEFNKALLAMYTNQAEQCRKISASLQSQSPEHLLPVLIQAAQLCREKQHTKAIELLQEFSDQHPENAAEIKLTMAQLKISQGNISKACLILRSIEELKHKPGMVSALVTMYSHEEDIDSAIEVFTQAIQWYQNHQPKSPAHLSLIREAANFKLKYGRKKEAISDLQQLWKQNPKDIHTLAQLISAYSLVDPEKAKALSKHLPSSDSMSLKVDVEALENSAGATYIRKKGGKVTGDSQPKEQGQGDLKKKKKKKKGKLPKNYDPKVTPDPERWLPMRERSYYRGRKKGKKKDQIGKGTQGATAGASSELDASKTVSSPPTSPRPGSAATVSASTSNIIPPRHQKPAGAPATKKKQQQKKKKGGKGGW srp68-srp72-2 4110128 N/A 4110132 unspecified role unspecified role neutral component neutral component purified purified region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 4110133 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 5 srp68-srp72-4 4110128 N/A 4110132 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal thioredoxin tag fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 4110133 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 7 Separation of a mixture of polypeptide 68c with fragments generated by mild digestion of 72b′ with chymotrypsin followed by analysis of the sucrose gradient fractions by SDS-PAGE. srp68-srp72-3 4110129 N/A 4110132 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal thioredoxin tag fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 4110133 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 6A srp72-srp68-1 4110130 N/A 4110132 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal thioredoxin tag fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 4110133 unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 6B srp72-srp68-2 4110131 N/A 4110132 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal thioredoxin tag fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 4110133 unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false 4110131