DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro mst microscale thermophoresis mst predetermined predetermined participant predetermined Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! cas9_strp1 CRISPR-associated endonuclease Cas9/Csn1 cas9 csn1 SPy_1046 SpCas9 SpyCas9 protein protein strp1 Streptococcus pyogenes serotype M1 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD DIP protein Q99ZW2 original sequence version: 77 a0a247d711_lismn acrIIA4 G3O21_003035 APD94_15065 GON91_02470 UI29_15095 protein protein lismn Listeria monocytogenes MNINDLIREIKNKDYTVKLSGTDSNSITQLIIRVNNDGNEYVISESENESIVEKFISAFKNGWNQEYEDEEEFYNDMQTITLKSELN F44958822FEFF9A5 urs000080e253_strpy_grna URS000080E253 gRNA grna guide rna guide RNA grna strpy Streptococcus pyogenes GGAAAUUAGGUGCGCUUGGCGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUU a0a459ndx2_lismn acrIIA2 D4271_13320 D4D89_13255 B1N40_08540 B4X79_13415 E0I39_13150 FJU19_12475 G3R95_002503 AF006_07510 APD94_09090 D4C60_14490 D4D22_14105 D5M63_13385 E1V33_15495 protein protein lismn Listeria monocytogenes MTLTRAQKKYAEAMHEFINMVDDFEESTPDFAKEVLHDSDYVVITKNEKYAVALCSLSTDECEYDTNLYLDEKLVDYSTVDVNGVTYYINIVETNDIDDLEIATDEDEMKSGNQEIILKSELK 47F691CB7CD8F4E3 a0a247d711_lismn-cas9-1 1 7 weight by bromide molecular weight estimation by bromide staining weight by bromide experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 1 a0a247d711_lismn-cas9-2 3 6 predetermined predetermined participant predetermined structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 5 predetermined predetermined participant predetermined structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Figure 2, Extended Data Table 1 a0a394vx67_lismn-cas9-1 2 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 8 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 4d cas9-a0a247d711_lismn-2 4 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) physical association physical association Extended Data Figure 1 cas9-a0a247d711_lismn-1 4 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 1 a0a247d711_lismn-cas9-3 4 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified A0A247D711:p.Asn12Thr N12T mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain N T Tue Jun 30 17:34:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn12Thr' A0A247D711:p.Gly38Ala G38A mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain G A Tue Jun 30 19:05:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly38Ala' gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range A0A247D711:p.Asp23Arg D23R mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 16:48:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp23Arg' ecoli Escherichia coli (strain K12) 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (AcrIIA4 mutant) a0a247d711_lismn-cas9-4 4 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified A0A247D711:p.Asn36Tyr N36Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N Y Mon Apr 29 11:10:17 BST 2019 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn36Tyr' Tue Jun 30 17:04:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn36Tyr' gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range A0A247D711:p.Asn39Arg N39R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N R Tue Jun 30 20:03:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn39Arg' A0A247D711:p.Glu70Arg E70R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E R Tue Jun 30 16:05:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu70Arg' A0A247D711:p.Asp69Arg D69R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 15:42:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp69Arg' A0A247D711:p.[Asp14Arg;Glu40Arg] D14R, E40R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E R certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:27:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp14Arg;Glu40Arg]' A0A247D711:p.[Asn36Arg;Asp69Arg] N36R, D69R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N R certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:51:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn36Arg;Asp69Arg]' ecoli Escherichia coli (strain K12) 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (AcrIIA4 mutant) a0a247d711_lismn-cas9-5 4 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified Q99ZW2:p.[Arg976Tyr;Lys1200Tyr] R976Y, K1200Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Y certain certain sequence position certain certain certain sequence position certain K Y Tue Jun 30 19:31:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg976Tyr;Lys1200Tyr]' Q99ZW2:p.[Lys1200Tyr;Ser1216Arg] K1200Y,S1216R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K Y certain certain sequence position certain certain certain sequence position certain S R Tue Jun 30 17:47:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys1200Tyr;Ser1216Arg]' Q99ZW2:p.Thr1138Tyr T1138Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain T Y Tue Jun 30 18:48:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr1138Tyr' Q99ZW2:p.Asp1332Arg D1332R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:59:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1332Arg' Q99ZW2:p.Arg1335Tyr R1335Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Y Tue Jun 30 17:27:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1335Tyr' Q99ZW2:p.Ser1216Thr S1216T mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S T Tue Jun 30 18:35:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1216Thr' ecoli Escherichia coli (strain K12) 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (SpyCas9 mutant) a0a247d711_lismn-cas9-6 2 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 4d