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a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second <t>Strep-Tactin</t> column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.
Pre Equilibrated Gravity Flow Strep Tactin Xt Column, supplied by IBA Lifesciences, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second <t>Strep-Tactin</t> column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.
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a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second <t>Strep-Tactin</t> column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.
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a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second <t>Strep-Tactin</t> column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.
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a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second Strep-Tactin column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.

Journal: Nature Plants

Article Title: Stages of biomolecular condensate formation in pro-β-carboxysome assembly

doi: 10.1038/s41477-026-02227-6

Figure Lengend Snippet: a , Two-step purification of the SIIApN/CM-H 10 hetero-complexes. Top: Workflow of the two-step purification. Middle: Schematic of the bicistronic plasmids for recombinant expression of SIIApN/CM-H 10 and SIIApN/CM Nt -H 10 hetero-complexes. Bottom: SDS-PAGE analysis of the purified proteins. The numbering of the lanes corresponds to the numbering in the workflow. Note that two protein forms are produced by the gene ccmM-H 10 , the full-length protein CM-H 10 and CM Ct -H 10 . CM Ct -H 10 is produced via an internal ribosome binding site and is in the flowthrough of the second Strep-Tactin column as it has no binding affinity for ApN. Representative data from two independent experiments are shown (n = 2). b , AlphaFold 3 structural model of (ApN) 3 :CM hetero-complex. End views of (ApN) 3 :CM in ribbon representation, highlighting the N- and C-termini and the encapsulation peptide (EP) of ApN. ApN, residues 1-161; CM, residues 1-198. The color bar indicates the confidence scores of the AF3 model (pLDDT), with high confidence (blue) and low confidence (orange) transitioning through the color spectrum. c , End view of the hetero-complex of ApN (residues 1-114) and CM Nt (residues 16-182) in ribbon representation, highlighting the position of the N-terminal tail of ApN1 protomer in the middle of the tetramer.

Article Snippet: The mixture was loaded onto a pre-equilibrated gravity-flow Strep–Tactin XT column (IBA), washed with buffer C and eluted with buffer C/50 mM biotin.

Techniques: Purification, Recombinant, Expressing, SDS Page, Produced, Binding Assay, Encapsulation