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/ Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. J. Biol. Chem. (0021-9258) 2011 imex curation Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem.utoronto.ca 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions ecoli Escherichia coli (strain K12) crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. Cells were exposed to a cell permeable cross-linker 1,5-difluoro-2,4-dinitrobenzene before lysis. The cross-linked samples where used for the pull down assay dzamble@chem. utoronto.ca dzamble@chem.utoronto.ca Accepted 2012-OCT-01 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. J. Biol. Chem. (0021-9258) 2011 imex curation Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem.utoronto.ca 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions ecoli Escherichia coli (strain K12) crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. The cross-linked samples were used for the pull down assays dzamble@chem. utoronto.ca dzamble@chem.utoronto.ca Accepted 2012-OCT-01 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. J. Biol. Chem. (0021-9258) 2011 imex curation Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem.utoronto.ca 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions ecoli Escherichia coli (strain K12) pull down pull down western blot western blot Immuno blot Immuno blot Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem. utoronto.ca dzamble@chem.utoronto.ca Accepted 2012-OCT-01 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. J. Biol. Chem. (0021-9258) 2011 imex curation Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem.utoronto.ca 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions ecoli Escherichia coli (strain K12) pull down pull down sequence tag sequence tag identification sequence tag Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem. utoronto.ca dzamble@chem.utoronto.ca Accepted 2012-OCT-01 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. The pulldown experiments from DPABF cells were resolved on SDS-polyacrylamide gels, and stained with Coomassie Blue dye. To identify some of these co-eluting proteins, bands were cut from the gels, and in-gel tryptic digests were performed followed by LC-MS/MS J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. Protein interactions and localization of the Escherichia coli accessory protein HypA during nickel insertion to [NiFe] hydrogenase. J. Biol. Chem. (0021-9258) 2011 imex curation Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. dzamble@chem.utoronto.ca 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions ecoli Escherichia coli (strain K12) crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Chan Chung K.C., Zamble D.B. Hydrogenase activity in DPABF cells producing the C-terminal tagged HypA (HypAStr) is comparable with that from cells producing untagged HypA (Fig. 1A), demonstrating that HypAStr is functional in hydrogenase maturation. The cross-linked samples were used for the pull down assays. The pull down proteins were analysed with a native gel electrophoresis and a Western blot dzamble@chem. utoronto.ca dzamble@chem.utoronto.ca Accepted 2012-OCT-01 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation hypb_ecoli Hydrogenase maturation factor HypB hypB Hydrogenase isoenzymes nickel incorporation protein HypB JW2697 b2727 protein protein ecoli Escherichia coli (strain K12) MCTTCGCGEGNLYIEGDEHNPHSAFRSAPFAPAARPKMKITGIKAPEFTPSQTEEGDLHYGHGEAGTHAPGMSQRRMLEVEIDVLDKNNRLAERNRARFAARKQLVLNLVSSPGSGKTTLLTETLMRLKDSVPCAVIEGDQQTVNDAARIRATGTPAIQVNTGKGCHLDAQMIADAAPRLPLDDNGILFIENVGNLVCPASFDLGEKHKVAVLSVTEGEDKPLKYPHMFAAASLMLLNKVDLLPYLNFDVEKCIACAREVNPEIEIILISATSGEGMDQWLNWLETQRCA DIP protein P0AAN3 original sequence version: 30 5B3AEB59AA6915C3 hyce_ecoli Formate hydrogenlyase subunit 5 Hydrogenase-3 component E hycE hevE b2721 JW2691 protein protein ecoli Escherichia coli (strain K12) MSEEKLGQHYLAALNEAFPGVVLDHAWQTKDQLTVTVKVNYLPEVVEFLYYKQGGWLSVLFGNDERKLNGHYAVYYVLSMEKGTKCWITVRVEVDANKPEYPSVTPRVPAAVWGEREVRDMYGLIPVGLPDERRLVLPDDWPDELYPLRKDSMDYRQRPAPTTDAETYEFINELGDKKNNVVPIGPLHVTSDEPGHFRLFVDGENIIDADYRLFYVHRGMEKLAETRMGYNEVTFLSDRVCGICGFAHSTAYTTSVENAMGIQVPERAQMIRAILLEVERLHSHLLNLGLACHFTGFDSGFMQFFRVRETSMKMAEILTGARKTYGLNLIGGIRRDLLKDDMIQTRQLAQQMRREVQELVDVLLSTPNMEQRTVGIGRLDPEIARDFSNVGPMVRASGHARDTRADHPFVGYGLLPMEVHSEQGCDVISRLKVRINEVYTALNMIDYGLDNLPGGPLMVEGFTYIPHRFALGFAEAPRGDDIHWSMTGDNQKLYRWRCRAATYANWPTLRYMLRGNTVSDAPLIIGSLDPCYSCTDRMTVVDVRKKKSKVVPYKELERYSIERKNSPLK DIP protein P16431 original sequence version: null 38A2BCCB01B27463 fdhf_ecoli Formate dehydrogenase H Formate-hydrogen-lyase-linked, selenocysteine-containing polypeptide Formate dehydrogenase-H subunit alpha fdhF b4079 JW4040 protein protein ecoli Escherichia coli (strain K12) MKKVVTVCPYCASGCKINLVVDNGKIVRAEAAQGKTNQGTLCLKGYYGWDFINDTQILTPRLKTPMIRRQRGGKLEPVSWDEALNYVAERLSAIKEKYGPDAIQTTGSSRGTGNETNYVMQKFARAVIGTNNVDCCARVUHGPSVAGLHQSVGNGAMSNAINEIDNTDLVFVFGYNPADSHPIVANHVINAKRNGAKIIVCDPRKIETARIADMHIALKNGSNIALLNAMGHVIIEENLYDKAFVASRTEGFEEYRKIVEGYTPESVEDITGVSASEIRQAARMYAQAKSAAILWGMGVTQFYQGVETVRSLTSLAMLTGNLGKPHAGVNPVRGQNNVQGACDMGALPDTYPGYQYVKDPANREKFAKAWGVESLPAHTGYRISELPHRAAHGEVRAAYIMGEDPLQTDAELSAVRKAFEDLELVIVQDIFMTKTASAADVILPSTSWGEHEGVFTAADRGFQRFFKAVEPKWDLKTDWQIISEIATRMGYPMHYNNTQEIWDELRHLCPDFYGATYEKMGELGFIQWPCRDTSDADQGTSYLFKEKFDTPNGLAQFFTCDWVAPIDKLTDEYPMVLSTVREVGHYSCRSMTGNCAALAALADEPGYAQINTEDAKRLGIEDEALVWVHSRKGKIITRAQVSDRPNKGAIYMTYQWWIGACNELVTENLSPITKTPEYKYCAVRVEPIADQRAAEQYVIDEYNKLKTRLREAALA C7C86F6F704A142D gatz_ecoli D-tagatose-1,6-bisphosphate aldolase subunit GatZ gatZ b2095 JW2082 protein protein ecoli Escherichia coli (strain K12) MKTLIARHKAGEHIGICSVCSAHPLVIEAALAFDRNSTRKVLIEATSNQVNQFGGYTGMTPADFREFVFTIADKVGFARERIILGGDHLGPNCWQQENADAAMEKSVELVKEYVRAGFSKIHLDASMSCAGDPIPLAPETVAERAAVLCFAAESVATDCQREQLSYVIGTEVPVPGGEASAIQSVHITHVEDAANTLRTHQKAFIARGLTEALTRVIAIVVQPGVEFDHSNIIHYQPQEAQPLAQWIENTRMVYEAHSTDYQTRTAYWELVRDHFAILKVGPALTFALREAIFALAQIEQELIAPENRSGCLAVIEEVMLDEPQYWKKYYRTGFNDSLLDIRYSLSDRIRYYWPHSRIKNSVETMMVNLEGVDIPLGMISQYLPKQFERIQSGELSAIPHQLIMDKIYDVLRAYRYGCAE This sequence has been withdrawn from Uniprot. 258D25D832E42580 258D25D832E42580 adhe_ecoli Aldehyde-alcohol dehydrogenase adhE ana b1241 JW1228 protein protein ecoli Escherichia coli (strain K12) MAVTNVAELNALVERVKKAQREYASFTQEQVDKIFRAAALAAADARIPLAKMAVAESGMGIVEDKVIKNHFASEYIYNAYKDEKTCGVLSEDDTFGTITIAEPIGIICGIVPTTNPTSTAIFKSLISLKTRNAIIFSPHPRAKDATNKAADIVLQAAIAAGAPKDLIGWIDQPSVELSNALMHHPDINLILATGGPGMVKAAYSSGKPAIGVGAGNTPVVIDETADIKRAVASVLMSKTFDNGVICASEQSVVVVDSVYDAVRERFATHGGYLLQGKELKAVQDVILKNGALNAAIVGQPAYKIAELAGFSVPENTKILIGEVTVVDESEPFAHEKLSPTLAMYRAKDFEDAVEKAEKLVAMGGIGHTSCLYTDQDNQPARVSYFGQKMKTARILINTPASQGGIGDLYNFKLAPSLTLGCGSWGGNSISENVGPKHLINKKTVAKRAENMLWHKLPKSIYFRRGSLPIALDEVITDGHKRALIVTDRFLFNNGYADQITSVLKAAGVETEVFFEVEADPTLSIVRKGAELANSFKPDVIIALGGGSPMDAAKIMWVMYEHPETHFEELALRFMDIRKRIYKFPKMGVKAKMIAVTTTSGTGSEVTPFAVVTDDATGQKYPLADYALTPDMAIVDANLVMDMPKSLCAFGGLDAVTHAMEAYVSVLASEFSDGQALQALKLLKEYLPASYHEGSKNPVARERVHSAATIAGIAFANAFLGVCHSMAHKLGSQFHIPHGLANALLICNVIRYNANDNPTKQTAFSQYDRPQARRRYAEIADHLGLSAPGDRTAAKIEKLLAWLETLKAELGIPKSIREAGVQEADFLANVDKLSEDAFDDQCTGANPRYPLISELKQILLDTYYGRDYVEGETAAKKEAAPAKAEKKAKKSA DIP protein P0A9Q7 original sequence version: 43 75469F57419C8C79 ch60_ecoli Chaperonin GroEL groEL 60 kDa chaperonin Chaperonin-60 GroEL protein groL mopA JW4103 b4143 protein protein ecoli Escherichia coli (strain K12) MAAKDVKFGNDARVKMLRGVNVLADAVKVTLGPKGRNVVLDKSFGAPTITKDGVSVAREIELEDKFENMGAQMVKEVASKANDAAGDGTTTATVLAQAIITEGLKAVAAGMNPMDLKRGIDKAVTAAVEELKALSVPCSDSKAIAQVGTISANSDETVGKLIAEAMDKVGKEGVITVEDGTGLQDELDVVEGMQFDRGYLSPYFINKPETGAVELESPFILLADKKISNIREMLPVLEAVAKAGKPLLIIAEDVEGEALATLVVNTMRGIVKVAAVKAPGFGDRRKAMLQDIATLTGGTVISEEIGMELEKATLEDLGQAKRVVINKDTTTIIDGVGEEAAIQGRVAQIRQQIEEATSDYDREKLQERVAKLAGGVAVIKVGAATEVEMKEKKARVEDALHATRAAVEEGVVAGGGVALIRVASKLADLRGQNEDQNVGIKVALRAMEAPLRQIVLNCGEEPSVVANTVKGGDGNYGYNAATEEYGNMIDMGILDPTKVTRSALQYAASVAGLMITTECMVTDLPKNDAADLGAAGGMGGMGGMGGMM CD3A0FB505F74AD1 CD3A0FB505F74AD1 dnak_ecol6 Chaperone protein DnaK dnaK HSP70 Heat shock 70 kDa protein Heat shock protein 70 c0019 protein protein ecol6 Escherichia coli O6:H1 (strain CFT073 / ATCC 700928 / UPEC) MGKIIGIDLGTTNSCVAIMDGTTPRVLENAEGDRTTPSIIAYTQDGETLVGQPAKRQAVTNPQNTLFAIKRLIGRRFQDEEVQRDVSIMPFKIIAADNGDAWVEVKGQKMAPPQISAEVLKKMKKTAEDYLGEPVTEAVITVPAYFNDAQRQATKDAGRIAGLEVKRIINEPTAAALAYGLDKGTGNRTIAVYDLGGGTFDISIIEIDEVDGEKTFEVLATNGDTHLGGEDFDSRLINYLVEEFKKDQGIDLRNDPLAMQRLKEAAEKAKIELSSAQQTDVNLPYITADATGPKHMNIKVTRAKLESLVEDLVNRSIEPLKVALQDAGLSVSDIDDVILVGGQTRMPMVQKKVAEFFGKEPRKDVNPDEAVAIGAAVQGGVLTGDVKDVLLLDVTPLSLGIETMGGVMTTLIAKNTTIPTKHSQVFSTAEDNQSAVTIHVLQGERKRAADNKSLGQFNLDGINPAPRGMPQIEVTFDIDADGILHVSAKDKNSGKEQKITIKASSGLNEDEIQKMVRDAEANAEADRKFEELVQTRNQGDHLLHSTRKQVEEAGDKLPADDKTAIESALTALETALKGEDKAAIEAKMQELAQVSQKLMEIAQQQHAQQQTAGADASANNAKDDDVVDAEFEEVKDKK 5A8589B21D7CD9C1 hypa_ecoli Hydrogenase maturation factor HypA hypA b2726 JW2696 protein protein ecoli Escherichia coli (strain K12) MHEITLCQRALELIEQQAAKHGAKRVTGVWLKIGAFSCVETSSLAFCFDLVCRGSVAEGCKLHLEEQEAECWCETCQQYVTLLTQRVRRCPQCHGDMLQIVADDGLQIRRIEIDQE 57458941783073D4 kka1_ecolx Aminoglycoside 3'-phosphotransferase aphA1 nptI APH(3')-I Kanamycin kinase, type I Neomycin-kanamycin phosphotransferase type I protein protein ecolx Escherichia coli MSHIQRETSCSRPRLNSNLDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF 3B715D9BBE8031B8 slyd_ecoli FKBP-type peptidyl-prolyl cis-trans isomerase SlyD slyD b3349 Metallochaperone SlyD JW3311 Histidine-rich protein WHP Rotamase Sensitivity to lysis protein D protein protein ecoli Escherichia coli (strain K12) MKVAKDLVVSLAYQVRTEDGVLVDESPVSAPLDYLHGHGSLISGLETALEGHEVGDKFDVAVGANDAYGQYDENLVQRVPKDVFMGVDELQVGMRFLAETDQGPVPVEITAVEDDHVVVDGNHMLAGQNLKFNVEVVAIREATEEELAHGHVHGAHDHHHDHDHDGCCGGHGHDHGHEHGGEGCCGGKGNGGCGCH DIP protein P0A9K9 original sequence version: 42 46DCC7C7ECD61DBA hypa-hyce-3 5 7 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey physical association physical association supplemental Fig. S3 hyce-hypa-1 5 13 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role prey prey physical association physical association supplemental Fig. S3 hypa-hyce-4 3 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey association association Figure 1B HycE co-eluted with HypAStr in the unprocessed form (supplemental Fig. S1). hyce-hypa-2 3 13 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 15 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association supplemental Fig. S2 hyce-hypa-3 1 13 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 15 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey physical association physical association Figure 2A, Supplemental data Figure S2 The use of cross-linker resulted in detection of increased amounts of co-eluting nickel insertion proteins with HycEStr (supplemental Fig. S2) hypa-hyce-5 1 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range 15 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey physical association physical association Figure 2B hypa-hyce-6 2 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey physical association physical association supplemental data Table S1 hyce-hypa-4 1 13 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3A Pulldown assays with HycEStr were performed in DHP-B strain (no expression of hypB) and delta slyD strain (no expression of slyD) hypa-hyce-7 1 7 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 3 B Pulldown assays with HypAStr were performed in DHP-B strain (no expression of hypB) and delta slyD strain (no expression of slyD) hypa-hyce-8 2 7 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association supplemental data Table S1 The assay was performed in the DHP-B E.coli strain with no HypB production when HycEStr pulldown experiments were performed from the DPABF strain, HypB could not be detected by Western blot (data not shown) or by LC-MS/MS (supplemental Table S1), supporting a role for HypA in mediating complex formation between HypB and HycE. hypa-hypb 1 15 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey physical association physical association Figure 3C They used a bacterial strain containing gene deletions of three hydrogenase isoform large subunits (hyaB, hybC, hycE) hypa-hypb-1 2 15 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey physical association physical association Supplemental data Table S2 They used a bacterial strain containing gene deletions of three hydrogenase isoform large subunits (hyaB, hybC, hycE) hyce-hypa-5 1 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role prey prey physical association physical association Figure 5C The pulldown assay was performed in the bacterial strain HYD723 (delta nikA), which lacks the soluble periplasmic component of the NikABCDE transporter A marked decrease in the amount of HypA pulled down was observed in HYD723 compared with the MC4100 wild-type strain When extra nickel was added to the growth media of the HYD723 cells only a very slight increase in the amount of HypA pulled down with HycEStr was observed (Fig. 5C). hypa-hyce-9 1 7 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey physical association physical association supplemental data Figure S6 The amount of HypB and SlyD pulled down with HypAStr from HYD723 cell lysates was not dramatically affected The pulldown assay was performed in the bacterial strain HYD723 (delta nikA), which lacks the soluble periplasmic component of the NikABCDE transporter A marked decrease in the amount of HycE pulled down was observed in HYD723 compared with the MC4100 wild-type strain hyce-hypa-6 1 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation P0A700:p.His2Ala mutation mutation certain certain sequence position certain certain certain sequence position certain H A Wed May 02 16:28:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his2ala' Tue Jun 30 16:09:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His2Ala' physical association physical association supplemental data Figure S7 adhe-apha1-1 4 10 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 14 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 8 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 6 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 13 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 15 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 7 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation association association Figure 6A, Supplemental data Table S1, Figure 1 Due to the abundance of proteins in the HypAStr elution fractions, clear HypB, SlyD, and HycE bands were not observed in the Coomassie stained gel; however, tryptic digests of elution fractions followed by LC-MS/MS as well as Western blotting did confirm their presence (Fig. 1, and supplemental Table S1). hyce-dnak 4 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation 12 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation association association Figure 6C, Supplemental data table S1 hyce-grol 2 15 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation strep-tag ll strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 11 unspecified role unspecified role prey prey physical association physical association Supplemental data Table S2 They use the DHP-C strain for this assay