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/ Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Experimental eye research 2019 imex curation Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease cross linker: DSS jint@ustc.edu.cn,wang_qiwei@126.com Accepted 2019-OCT-29 AT 12:02 GMT AT 12:02 GMT by MGT Experimental eye research 2019 Only protein-protein interactions imex curation imex curation Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Experimental eye research 2019 imex curation Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com Accepted 2019-OCT-29 AT 12:02 GMT AT 12:02 GMT by MGT Experimental eye research 2019 Only protein-protein interactions imex curation imex curation Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Experimental eye research 2019 imex curation Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com Accepted 2019-OCT-29 AT 12:02 GMT AT 12:02 GMT by MGT Experimental eye research 2019 Only protein-protein interactions imex curation imex curation Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Experimental eye research 2019 imex curation Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com Accepted 2019-OCT-29 AT 12:02 GMT AT 12:02 GMT by MGT Experimental eye research 2019 Only protein-protein interactions imex curation imex curation Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Biochemical characterization of G64W mutant of acidic beta-crystallin 4. Experimental eye research 2019 imex curation Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fcs fluorescence correlation spectroscopy FCS FCS fcs tag fluorescence tag visualisation by fluorescence tag fluorescence Li W., Ji Q., Wei Z., Chen YL., Zhang Z., Yin X., Aghmiuni SK., Liu M., Chen W., Shi L., Chen Q., Du X., Yu L., Cao MJ., Wang Z., Huang S., Jin T., Wang Q. Rare diseases - Interactions investigated in the context of Rare genetic disease jint@ustc.edu.cn,wang_qiwei@126.com Accepted 2019-OCT-29 AT 12:02 GMT AT 12:02 GMT by MGT Experimental eye research 2019 Only protein-protein interactions imex curation imex curation cryaa_human Alpha-crystallin A chain CRYAA CRYA1 HSPB4 Heat shock protein beta-4 protein protein human Homo sapiens Human MDVTIQHPWFKRTLGPFYPSRLFDQFFGEGLFEYDLLPFLSSTISPYYRQSLFRTVLDSGISEVRSDRDKFVIFLDVKHFSPEDLTVKVQDDFVEIHGKHNERQDDHGYISREFHRRYRLPSNVDQSALSCSLSADGMLTFCGPKIQTGLDATHAERAIPVSREEKPTSAPSS DIP protein P02489 original sequence version: 121 DIP protein P02489 original sequence version: 131 81804A8439837D50 crbb1_human Beta-crystallin B1 CRYBB1 Beta-B1 crystallin protein protein human Homo sapiens Human MSQAAKASASATVAVNPGPDTKGKGAPPAGTSPSPGTTLAPTTVPITSAKAAELPPGNYRLVVFELENFQGRRAEFSGECSNLADRGFDRVRSIIVSAGPWVAFEQSNFRGEMFILEKGEYPRWNTWSSSYRSDRLMSFRPIKMDAQEHKISLFEGANFKGNTIEIQGDDAPSLWVYGFSDRVGSVKVSSGTWVGYQYPGYRGYQYLLEPGDFRHWNEWGAFQPQMQSLRRLRDKQWHLEGSFPVLATEPPK Defects in CRYBB1 are the cause of autosomal recessive congenital nuclear cataract type 3 (CATCN3) [MIM:611544]. CATCN3 is a form of non-syndromic congenital cataract. Non-syndromic congenital cataracts vary markedly in severity and morphology, affecting the nuclear, cortical, polar, or subcapsular parts of the lens or, in severe cases, the entire lens, with a variety of types of opacity. They are one of the major causes of vision loss in children worldwide and are responsible for approximately one third of blindness in infants. Congenital cataracts can lead to permanent blindness by interfering with the sharp focus of light on the retina during critical developmental intervals mint Crystallins are the dominant structural components of the vertebrate eye lens 93D81A9C95A86F7F crba4_human Beta-crystallin A4 CRYBA4 Beta-A4 crystallin protein protein human Homo sapiens Human MTLQCTKSAGPWKMVVWDEDGFQGRRHEFTAECPSVLELGFETVRSLKVLSGAWVGFEHAGFQGQQYILERGEYPSWDAWGGNTAYPAERLTSFRPAACANHRDSRLTIFEQENFLGKKGELSDDYPSLQAMGWEGNEVGSFHVHSGAWVCSQFPGYRGFQYVLECDHHSGDYKHFREWGSHAPTFQVQSIRRIQQ Crystallins do not turn over as the lens ages, providing ample opportunity for post-translational modifications or oxidations. These modifications may change crystallin solubility properties and favor senile cataract mint Crystallins are the dominant structural components of the vertebrate eye lens Defects in CRYBA4 are the cause of lamellar cataract 2 [MIM:610425]. Cataracts are a leading cause of blindness worldwide, affecting all societies. A significant proportion of cases are genetically determined. More than 15 genes for cataracts have been identified, of which the crystallin genes are the most commonly mutated. Lamellar cataract 2 is an autosomal dominant congenital cataract Defects in CRYBA4 are a cause of isolated microphthalmia with cataract 4 (MCOPCT4) [MIM:610426]. Microphtalmia consists of a development defect causing moderate or severe reduction in size of the eye. Opacities of the cornea and lens, scaring of the retina and choroid, and other abnormalities like cataract may also be present 104EC68E3CDE5740 cryba4-cryba4-1 2 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 20:23:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 18:29:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' physical association physical association Fig. 4E,F cryba4-cryba4-2 3 8 unspecified role unspecified role neutral component neutral component purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range p.Gly64Trp mutation with complex effect mutation with complex effect certain certain sequence position certain certain certain sequence position certain G W physical association physical association Fig. 4B The elution volume of CRYBA4WT-MBP fusion protein was around 15 ml, which corresponding to a molecular weight of about 150 kDa (Fig. 4C), suggesting it is a dimer in solution. Nevertheless, the elution volume of CRYBA4G64W-MBP fusion protein was near 9 ml, which corresponding to a molecular weight of more than 670 kDa (Fig. 4C), indicating the formation of large protein aggregates. cryba4-1 3 8 unspecified role unspecified role neutral component neutral component purified purified P53673:p.Gly64Trp mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 17:03:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 unspecified role unspecified role neutral component neutral component purified purified P53673:p.Gly64Trp mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 17:37:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 4B The elution volume of CRYBA4WT-MBP fusion protein was around 15 ml, which corresponding to a molecular weight of about 150 kDa (Fig. 4C), suggesting it is a dimer in solution. Nevertheless, the elution volume of CRYBA4G64W-MBP fusion protein was near 9 ml, which corresponding to a molecular weight of more than 670 kDa (Fig. 4C), indicating the formation of large protein aggregates. cryba4-cryba4-3 1 8 unspecified role unspecified role neutral component neutral component purified purified P53673:p.Gly64Trp mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 19:42:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp 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 purified purified P53673:p.Gly64Trp mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 18:24:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp 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. 4D Crosslink experiment also showed that the CRYBA4WT-MBP presented mainly as dimers, while the MBP-mutant existed as aggregates (Fig. 4D). cryba4-6 1 8 unspecified role unspecified role neutral component neutral component purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range p.Gly64Trp mutation with complex effect mutation with complex effect certain certain sequence position certain certain certain sequence position certain G W ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 4D Crosslink experiment also showed that the CRYBA4WT-MBP presented mainly as dimers, while the MBP-mutant existed as aggregates (Fig. 4D). cryba4-cryba4-4 5 8 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 17:32:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 16:52:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 4G, H These results indicate that the G64W mutant has a tendency of forming larger fluorescent particles compared to its WT counterpart (Fig. 4H). cryba4-cryba4-5 2 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 17:37:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' physical association physical association Fig. 6A crybb1-cryba4-1 2 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed P53673:p.Gly64Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 16:21:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Fig. 7A cryba4-cryba4-6 4 7 unspecified role unspecified role bait bait purified purified strep tag strep tag strep tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey purified purified his tag 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 P53673:p.Gly64Trp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain G W Tue Jun 30 17:51:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly64Trp' mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp 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