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/ Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Journal of Lipid Research 2011 imex curation Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. robert.kiss@mcgill.ca 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro crosslink cross-linking study crosslink weight identificat confirmation by molecular weight weight identificat Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. cross-linker: dimethyl suberimidate (DMS) mutation Ser36Ala was remapped to Ser60Ala, matching to the following authors' statement:"Interestingly, there have been very few reported mutations in this linker region: 34STOP (functional), A37T (variant-nonfunctional), G39E (variant-nonfunctional) (taken from the Human Gene Mutation Database; www.hgmd.cf.ac.uk) (19). We have recently identified a novel heterozygous mutation (S36A) in a 63-year-old male with HDL deficiency " robert.kiss@mcgill.ca Accepted 2019-DEC-02 AT 11:01 GMT AT 11:01 GMT by LUANA Journal of Lipid Research 2011 Only protein-protein interactions imex curation imex curation Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Journal of Lipid Research 2011 imex curation Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. robert.kiss@mcgill.ca 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by comassie molecular weight estimation by coomasie staining weight by comassie Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. mutation Ser36Ala was remapped to Ser60Ala, matching to the following authors' statement:"Interestingly, there have been very few reported mutations in this linker region: 34STOP (functional), A37T (variant-nonfunctional), G39E (variant-nonfunctional) (taken from the Human Gene Mutation Database; www.hgmd.cf.ac.uk) (19). We have recently identified a novel heterozygous mutation (S36A) in a 63-year-old male with HDL deficiency " robert.kiss@mcgill.ca Accepted 2019-DEC-02 AT 11:01 GMT AT 11:01 GMT by LUANA Journal of Lipid Research 2011 Only protein-protein interactions imex curation imex curation Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation. Journal of Lipid Research 2011 imex curation Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. robert.kiss@mcgill.ca 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Weers PM., Patel AB., Wan LC., Guigard E., Kay CM., Hafiane A., McPherson R., Marcel YL., Kiss RS. mutation Ser36Ala was remapped to Ser60Ala, matching to the following authors' statement:"Interestingly, there have been very few reported mutations in this linker region: 34STOP (functional), A37T (variant-nonfunctional), G39E (variant-nonfunctional) (taken from the Human Gene Mutation Database; www.hgmd.cf.ac.uk) (19). We have recently identified a novel heterozygous mutation (S36A) in a 63-year-old male with HDL deficiency " robert.kiss@mcgill.ca Accepted 2019-DEC-02 AT 11:01 GMT AT 11:01 GMT by LUANA Journal of Lipid Research 2011 Only protein-protein interactions imex curation imex curation apoa1_human Apolipoprotein A-I APOA1 Apolipoprotein A1 protein protein human Homo sapiens Human MKAAVLTLAVLFLTGSQARHFWQQDEPPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ DIP protein P02647 original sequence version: 116 DIP protein P02647 original sequence version: 125 DIP protein P02647 original sequence version: 147 DIP protein P02647 original sequence version: 162 1A28B8366E620310 apo1_dimer-1 1 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S A 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.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 7B ApoA-I was then crosslinked with DMS and analyzed by SDS-PAGE. This revealed that the majority of the S36A protein was in a monomeric state. However, WT apoA-I showed multiple cross-linked species of dimer, tetramer, and the appearance of a prominent band in the top portion of the gel representing apoA-I octomers (Fig. 7B). apoa1-15 2 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 18:29:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 7A To gain insight into the oligomerization state of S36A apoA-I, the proteins were analyzed by nondenaturing-PAGE. The gel showed that WT apoA-I was present as multiple bands, indicative of protein-protein interactions resulting in the formation of dimers and tetramers (Fig. 7A). The migration pattern of an identical concentration of S36A apoA-I was distinctively different, showing a more homogeneous apoA-I population. apoa1-16 1 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 19:01:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 7B ApoA-I was then crosslinked with DMS and analyzed by SDS-PAGE. This revealed that the majority of the S36A protein was in a monomeric state. However, WT apoA-I showed multiple cross-linked species of dimer, tetramer, and the appearance of a prominent band in the top portion of the gel representing apoA-I octomers (Fig. 7B). apoa1-17 1 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 18:07:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 7B ApoA-I was then crosslinked with DMS and analyzed by SDS-PAGE. This revealed that the majority of the S36A protein was in a monomeric state. However, WT apoA-I showed multiple cross-linked species of dimer, tetramer, and the appearance of a prominent band in the top portion of the gel representing apoA-I octomers (Fig. 7B). apoa1-18 3 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 17:52:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 8 The modeling of both systems showed evidence of monomer-tetramer with the monomer being predominant, more so in the case of the S36A apoA-I. As well, the calculated association constant (Ka) values were 8.561 × 1014 M−1for WT apoA-I and 6.119 × 1014 M−1for S36A apoA-I (Fig. 8). The global molecular average results suggested that S36A apoA-I clearly had less aggregation than WT apoA-I. apoa1-14 2 4 unspecified role unspecified role neutral component neutral component purified purified P02647:p.Ser60Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 18:49:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser60Ala' human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction Fig. 7A To gain insight into the oligomerization state of S36A apoA-I, the proteins were analyzed by nondenaturing-PAGE. The gel showed that WT apoA-I was present as multiple bands, indicative of protein-protein interactions resulting in the formation of dimers and tetramers (Fig. 7A). The migration pattern of an identical concentration of S36A apoA-I was distinctively different, showing a more homogeneous apoA-I population.