MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. FEBS Lett. (0014-5793) 2013 imex curation Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus Only protein-protein interactions imex curation in vitro In vitro fluorescence fluorescence technology fluorescence experimental particp experimental participant identification experimental particp Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus FEBS Lett. (0014-5793) 2013 IMEx Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. FEBS Lett. (0014-5793) 2013 imex curation Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus Only protein-protein interactions imex curation in vitro In vitro crosslink cross-linking study crosslink weight by comassie molecular weight estimation by coomasie staining weight by comassie Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus FEBS Lett. (0014-5793) 2013 IMEx Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein. FEBS Lett. (0014-5793) 2013 imex curation Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Lo YS., Lin SY., Wang SM., Wang CT., Chiu YL., Huang TH., Hou MH. Coronavirus - Interactions investigated in the context of Coronavirus FEBS Lett. (0014-5793) 2013 IMEx ncap_cvh22 Nucleoprotein N Nucleocapsid protein 7 protein protein hcov-229e Human coronavirus 229E HCoV-229E MATVKWADASEPQRGRQGRIPYSLYSPLLVDSEQPWKVIPRNLVPINKKDKNKLIGYWNVQKRFRTRKGKRVDLSPKLHFYYLGTGPHKDAKFRERVEGVVWVAVDGAKTEPTGYGVRRKNSEPEIPHFNQKLPNGVTVVEEPDSRAPSRSQSRSQSRGRGESKPQSRNPSSDRNHNSQDDIMKAVAAALKSLGFDKPQEKDKKSAKTGTPKPSRNQSPASSQTSAKSLARSQSSETKEQKHEMQKPRWKRQPNDDVTSNVTQCFGPRDLDHNFGSAGVVANGVKAKGYPQFAELVPSTAAMLFDSHIVSKESGNTVVLTFTTRVTVPKDHPHLGKFLEELNAFTREMQQHPLLNPSALEFNPSQTSPATAEPVRDEVSIETDIIDEVN mint homomint D3D86CFE10EEDCA1 n-23 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction f2b mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-21 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction f2a mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-24 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction f2c mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-25 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 4.0 direct interaction direct interaction f2c mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-26 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 4.0 direct interaction direct interaction f2a mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-22 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 4.0 direct interaction direct interaction f2b mint The truncated N245-350 protein predominantly displays a dimeric quaternary structure with a small amount of tetramers (Figure 2A), exhibiting a Kd,24 value of 256 μM. With an extended C-terminal tail that includes residue 370 or residue 376, the respective truncated N245-370 and N245-376 proteins also exist as dimers in solution with the major peaks exhibiting Kd,24 values of 177 and 159 μM, respectively (Figure 2B and 2C). n-20 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction f3a mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. n-27 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 4.0 direct interaction direct interaction f3a mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. n-28 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 8.0 direct interaction direct interaction f3a mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. n-29 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 2.0 direct interaction direct interaction mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. f3b f6 Figure 6 shows the distribution plots of the N245-389 protein in the absence and presence of the C-terminal tail peptide. In the absence of the C-terminal tail peptide, N245-389 formed a stable dimer, tetramer, and octamer with S-values of approximately 2.85, 5.30 and 7.75, respectively, corresponding to the molecular masses of 31, 69, and 131 kDa, respectively. When the concentrations of the C-terminal tail peptide were increased, the N245-389 tetramer and octamer peak decreased, whereas the N245-389 dimer peak increased (Figure 6). When the molar ratio of N245-389/N377-389 was 0.25 and 0.1, the Kd,24 value for N245-389 was approximately 6.1 μM and 9.7 μM, respectively, and it is significantly higher than that of N245-389 in the absence of the C-terminal tail peptide. These results indicate that the N245-389 tetramer and octamer was significantly dissociated into dimers in the presence of the peptide (N377-389) and suggest that the C-terminal tail peptide may complete the tetramer interface of N245-389 and interfere with the oligomerization of N245-389. n-30 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 4.0 direct interaction direct interaction mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. Figure 6 shows the distribution plots of the N245-389 protein in the absence and presence of the C-terminal tail peptide. In the absence of the C-terminal tail peptide, N245-389 formed a stable dimer, tetramer, and octamer with S-values of approximately 2.85, 5.30 and 7.75, respectively, corresponding to the molecular masses of 31, 69, and 131 kDa, respectively. When the concentrations of the C-terminal tail peptide were increased, the N245-389 tetramer and octamer peak decreased, whereas the N245-389 dimer peak increased (Figure 6). When the molar ratio of N245-389/N377-389 was 0.25 and 0.1, the Kd,24 value for N245-389 was approximately 6.1 μM 9 and 9.7 μM, respectively, and it is significantly higher than that of N245-389 in the absence of the C-terminal tail peptide. These results indicate that the N245-389 tetramer and octamer was significantly dissociated into dimers in the presence of the peptide (N377-389) and suggest that the C-terminal tail peptide may complete the tetramer interface of N245-389 and interfere with the oligomerization of N245-389. f3b f6 n-31 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 8.0 direct interaction direct interaction mint With an extended C-terminal tail that includes residue 382 or residue 389, the respective truncated N245-382 and N245-389 proteins demonstrated significant shifts in the equilibrium from dimers to tetramers and octamers, with significant decreases in the Kd,24 values (3.83 and 3.50 μM, respectively) (Figure 3A and 3B). These results indicate that residues 377-389 at the end of the C terminus are necessary for the oligomerization of the HCoV-229 N protein. Figure 6 shows the distribution plots of the N245-389 protein in the absence and presence of the C-terminal tail peptide. In the absence of the C-terminal tail peptide, N245-389 formed a stable dimer, tetramer, and octamer with S-values of approximately 2.85, 5.30 and 7.75, respectively, corresponding to the molecular masses of 31, 69, and 131 kDa, respectively. When the concentrations of the C-terminal tail peptide were increased, the N245-389 tetramer and octamer peak decreased, whereas the N245-389 dimer peak increased (Figure 6). When the molar ratio of N245-389/N377-389 was 0.25 and 0.1, the Kd,24 value for N245-389 was approximately 6.1 μM 9 and 9.7 μM, respectively, and it is significantly higher than that of N245-389 in the absence of the C-terminal tail peptide. These results indicate that the N245-389 tetramer and octamer was significantly dissociated into dimers in the presence of the peptide (N377-389) and suggest that the C-terminal tail peptide may complete the tetramer interface of N245-389 and interfere with the oligomerization of N245-389. f3b f6 n-n-17 3 N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined C-terminal tail peptide 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f6 homomint mint Figure 6 shows the distribution plots of the N245-389 protein in the absence and presence of the C-terminal tail peptide. In the absence of the C-terminal tail peptide, N245-389 formed a stable dimer, tetramer, and octamer with S-values of approximately 2.85, 5.30 and 7.75, respectively, corresponding to the molecular masses of 31, 69, and 131 kDa, respectively. When the concentrations of the C-terminal tail peptide were increased, the N245-389 tetramer and octamer peak decreased, whereas the N245-389 dimer peak increased (Figure 6). When the molar ratio of N245-389/N377-389 was 0.25 and 0.1, the Kd,24 value for N245-389 was approximately 6.1 μM 9 and 9.7 μM, respectively, and it is significantly higher than that of N245-389 in the absence of the C-terminal tail peptide. These results indicate that the N245-389 tetramer and octamer was significantly dissociated into dimers in the presence of the peptide (N377-389) and suggest that the C-terminal tail peptide may complete the tetramer interface of N245-389 and interfere with the oligomerization of N245-389. n-n-18 1 C-terminal tail peptide 4 peptide synthesis peptide synthesis 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain N 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f5 homomint mint To explore whether a C-terminal tail peptide (residues 377-389) can compete with the oligomerization site of the C-terminal domain of the HCoV-229E N protein and interfere with its oligomerization, we synthesized a C-terminal tail peptide (N377-389) and characterized the binding between N245-389 and this C-terminal tail peptide. First, we utilized fluorescence to monitor the protein/peptide binding because N245-389 contains one tryptophan residue, which contributes to its intrinsic fluorescence. The fluorescence emission spectra for N245-389 showed the maximal emission wavelength at approximately 332 nm with a fluorescence intensity of 142.4 AU (Figure 5A). At the C-terminal tail peptide concentrations of 5, 10, 20, 50, 75 and 100 μM, the C-terminal tail peptide decreased the fluorescence intensity of N245-389 at approximately 332 nm by 8.7, 19, 23.5, 25.4, 26.9 and 28.9 AU, respectively, after 4 hours of peptide addition. The N245-389 fluorescence decreased with increasing concentrations of the C-terminal tail peptide, which suggests that this decrease reflected the interaction of N245-389 with the C-terminal tail peptide. As shown in Figure 5B, the fluorescence quenching of N245-389 by the C-terminal tail peptide was analyzed with a Hill plot after the addition of the peptide. The dissociation constant of the C-terminal tail peptide for N245-389 was calculated to be 7.43 μM. n-32 2 N 4 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 his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction sf3a mint A crosslinking assay was also conducted to 10 investigate oligomerization by the C-terminal domain of the N protein. As shown in Figure S3A, the crosslinking studies of N245-350 also detected dimers and tetramers. n-33 2 N 4 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 his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 4.0 direct interaction direct interaction sf3a mint A crosslinking assay was also conducted to 10 investigate oligomerization by the C-terminal domain of the N protein. As shown in Figure S3A, the crosslinking studies of N245-350 also detected dimers and tetramers. n-34 2 N 4 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 2.0 direct interaction direct interaction sf3b mint Compared to N245-350, the crosslinking results showed that N245-389 forms dimers and tetramers as well as high-order oligomers (Figure S3B).. n-35 2 N 4 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 his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 4.0 direct interaction direct interaction sf3b mint Compared to N245-350, the crosslinking results showed that N245-389 forms dimers and tetramers as well as high-order oligomers (Figure S3B).. n-n-19 2 N 4 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 his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain N 4 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction sf3b homomint mint Compared to N245-350, the crosslinking results showed that N245-389 forms dimers and tetramers as well as high-order oligomers (Figure S3B)..