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/ Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mol. Cell (1097-2765) 2006 imex curation Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp Accepted 2014-JAN-20 by MGT Mol. Cell (1097-2765) 2006 Only protein-protein interactions imex curation Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mol. Cell (1097-2765) 2006 imex curation Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro sulfertransferase sulfurtransferase assay sulfertransferase tag visualisation tag visualisation Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp Accepted 2014-JAN-20 by MGT Mol. Cell (1097-2765) 2006 Only protein-protein interactions imex curation Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mechanistic insights into sulfur relay by multiple sulfur mediators involved in thiouridine biosynthesis at tRNA wobble positions. Mol. Cell (1097-2765) 2006 imex curation Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight by bromide molecular weight estimation by bromide staining weight by bromide Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T. Sulfur is transferred from [35S]cysteine to either IscS, MnmA or tRNA(Glu). The detection is by liquid scintillation and thiolation of tRNA was also visualised on a phosphor imaging plate. ts@chembio.t.u-tokyo.ac.jp Accepted 2014-JAN-20 by MGT Mol. Cell (1097-2765) 2006 Only protein-protein interactions imex curation tusa_ecoli Sulfur carrier protein TusA tRNA 2-thiouridine synthesizing protein A tusA b3470 sirA yhhP JW3435 Sulfur mediator TusA Sulfur transfer protein TusA protein protein ecoli Escherichia coli (strain K12) MTDLFSSPDHTLDALGLRCPEPVMMVRKTVRNMQPGETLLIIADDPATTRDIPGFCTFMEHELVAKETDGLPYRYLIRKGG 63AA7DE4816A3274 pyridoxal 5'-phosphate pyridoxal 5'-phosphate 3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]-4-pyridinecarboxaldehyde 3-hydroxy-5-(hydroxymethyl)-2-methylisonicotinaldehyde 5-phosphate codecarboxylase Phosphoric acid mono-(4-formyl-5-hydroxy-6-methyl-pyridin-3-ylmethyl) ester pyridoxal 5-monophosphoric acid ester pyridoxal 5'-(dihydrogen phosphate) Pyridoxal 5'-phosphate Pyridoxal 5-phosphate Pyridoxal phosphate PYRIDOXAL-5'-PHOSPHATE (4-formyl-5-hydroxy-6-methylpyridin-3-yl)methyl dihydrogen phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) NGVDGCNFYWLIFO-UHFFFAOYSA-N NGVDGCNFYWLIFO-UHFFFAOYSA-N tusb_ecoli Protein TusB tusB yheL tRNA 2-thiouridine synthesizing protein B b3343 JW3305 protein protein ecoli Escherichia coli (strain K12) MLHTLHRSPWLTDFAALLRLLSEGDELLLLQDGVTAAVDGNRYLESLRNAPIKVYALNEDLIARGLTGQISNDIILIDYTDFVRLTVKHPSQMAW 0EEF7560CCB4D608 0EEF7560CCB4D608 mnma_ecoli tRNA-specific 2-thiouridylase MnmA mnmA asuE trmU b1133 ycfB JW1119 protein protein ecoli Escherichia coli (strain K12) MSETAKKVIVGMSGGVDSSVSAWLLQQQGYQVEGLFMKNWEEDDGEEYCTAAADLADAQAVCDKLGIELHTVNFAAEYWDNVFELFLAEYKAGRTPNPDILCNKEIKFKAFLEFAAEDLGADYIATGHYVRRADVDGKSRLLRGLDSNKDQSYFLYTLSHEQIAQSLFPVGELEKPQVRKIAEDLGLVTAKKKDSTGICFIGERKFREFLGRYLPAQPGKIITVDGDEIGEHQGLMYHTLGQRKGLGIGGTKEGTEEPWYVVDKDVENNILVVAQGHEHPRLMSVGLIAQQLHWVDREPFTGTMRCTVKTRYRQTDIPCTVKALDDDRIEVIFDEPVAAVTPGQSAVFYNGEVCLGGGIIEQRLPLPV DIP protein P25745 original sequence version: null 22CA92676D0805CC tuse_ecoli Sulfurtransferase TusE tRNA 2-thiouridine synthesizing protein E tusE yccK b0969 JW0952 protein protein ecoli Escherichia coli (strain K12) MLIFEGKEIETDTEGYLKESSQWSEPLAVVIAENEGISLSPEHWEVVRFVRDFYLEFNTSPAIRMLVKAMANKFGEEKGNSRYLYRLFPKGPAKQATKIAGLPKPVKCI FDB0CFF89114EBE7 iscs_ecoli Cysteine desulfurase IscS iscS b2530 JW2514 yfhO yzzO ThiI transpersulfidase NifS protein homolog TusA transpersulfidase nuvC protein protein ecoli Escherichia coli (strain K12) MKLPIYLDYSATTPVDPRVAEKMMQFMTMDGTFGNPASRSHRFGWQAEEAVDIARNQIADLVGADPREIVFTSGATESDNLAIKGAANFYQKKGKHIITSKTEHKAVLDTCRQLEREGFEVTYLAPQRNGIIDLKELEAAMRDDTILVSIMHVNNEIGVVQDIAAIGEMCRARGIIYHVDATQSVGKLPIDLSQLKVDLMSFSGHKIYGPKGIGALYVRRKPRVRIEAQMHGGGHERGMRSGTLPVHQIVGMGEAYRIAKEEMATEMERLRGLRNRLWNGIKDIEEVYLNGDLEHGAPNILNVSFNYVEGESLIMALKDLAVSSGSACTSASLEPSYVLRALGLNDELAHSSIRFSLGRFTTEEEIDYTIELVRKSIGRLRDLSPLWEMYKQGVDLNSIEWAHH 1E4F2E6F9CD266B0 1E4F2E6F9CD266B0 tusc_ecoli Protein TusC tusC b3344 tRNA 2-thiouridine synthesizing protein C yheM JW3306 protein protein ecoli Escherichia coli (strain K12) MKRIAFVFSTAPHGTAAGREGLDALLATSALTDDLAVFFIADGVFQLLPGQKPDAVLARDYIATFKLLGLYDIEQCWVCAASLRERGLDPQTPFVVEATPLEADALRRELANYDVILRF 2AF593ADC435CA0D 2AF593ADC435CA0D cysteine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) tusd_ecoli Sulfurtransferase TusD tusD yheN tRNA 2-thiouridine synthesizing protein D b3345 JW3307 protein protein ecoli Escherichia coli (strain K12) MRFAIVVTGPAYGTQQASSAFQFAQALIADGHELSSVFFYREGVYNANQLTSPASDEFDLVRAWQQLNAQHGVALNICVAAALRRGVVDETEAGRLGLASSNLQQGFTLSGLGALAEASLTCDRVVQF 87B2754849A0FEFF 87B2754849A0FEFF trnaglu CHEBI:29175 trna transfer rna transfer ribonucleic acid tRNA transfer RNA trna yeast Saccharomyces cerevisiae Baker's yeast iscs-mnma-1 2 13 unspecified role unspecified role neutral component neutral component u8c mutation mutation certain certain sequence position certain certain certain sequence position certain u34a mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 7 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 ecoli Escherichia coli (strain K12) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 4a, b The thiolation of tRNA(Glu) is enhanced by the addition of TusA (P0A890), TusA and TusBCD complex (P45530, P45531, P45532), TusA and TusE (P0AB18) and - rather significantly - enhanced by the addition of TusA, TusBCD and TusE. IscS and MnmA are the minimum essential factors for 2-thiolation of mnm5s2U; addition of TusA alone enhances the interaction, and addition of the TusBCD complex and/or TusE stimulates 2-thiolation activity further. iscs-tusade-1 2 4 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role neutral component neutral component u8c mutation mutation certain certain sequence position certain certain certain sequence position certain u34a mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 12 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 ecoli Escherichia coli (strain K12) 8 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 ecoli Escherichia coli (strain K12) 5 cofactor cofactor coenzyme neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 4c In addition to the 2-thiouridine of the tRNAGlu(U8C), IscS, TusA, TusD, and TusE were clearly labeled by [35S]. These labeled protein bands disappeared when treated with reducing agent (DTT). iscs-tusa-1 1 9 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 ecoli Escherichia coli (strain K12) 4 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5a iscs-tusa-2 2 5 cofactor cofactor coenzyme neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 4 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 P0A890:p.Cys19Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:53:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys19ser' Tue Jun 30 19:46:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys19Ser' ecoli Escherichia coli (strain K12) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 5b, d trnaglu-mnma-1 3 7 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role bait bait u8c mutation mutation certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 6a trnaglu-mnma-2 3 13 unspecified role unspecified role bait bait u8c mutation mutation certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 6a tusbcd-tuse 1 8 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 ecoli Escherichia coli (strain K12) 12 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. 6b Figure legends and text references for Figures 6b and 6c have been inverted! mnma-tuse 1 7 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 ecoli Escherichia coli (strain K12) 8 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 6c Figure legends and text references for Figures 6b and 6c have been inverted! iscs-tusade-2 2 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 8 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 ecoli Escherichia coli (strain K12) 4 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 ecoli Escherichia coli (strain K12) 12 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 ecoli Escherichia coli (strain K12) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 6d lane 7 iscs-tusad-1 2 4 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 ecoli Escherichia coli (strain K12) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 12 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 ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 6d lane 5 iscs-tusa-3 2 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 4 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 ecoli Escherichia coli (strain K12) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 6d lane 2 iscs-cysteine-1 2 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 6d lane 1 iscs-tusae-1 2 8 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 ecoli Escherichia coli (strain K12) 9 enzyme enzyme 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 ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 4 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 ecoli Escherichia coli (strain K12) sulfurtransfer sulfurtransfer reaction sulfurtransfer Fig. 6d lane 6