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. 2018 Jun 11;19(7):e45543. doi: 10.15252/embr.201745543

Figure EV1. Cas1:Cas2‐DnaQ composition analysis.

Figure EV1

Cas1 StrepII‐tagged at N‐terminus (S‐Cas1) was co‐expressed with either full‐length Cas2‐DnaQ protein, Cas2 domain, or DnaQ domain tagged with His6‐tag at the C‐terminus (Cas2‐DnaQ‐H, Cas2‐H, and DnaQ‐H, respectively). Co‐elution of S‐Cas1 with Cas2‐DnaQ‐H or separate Cas2‐H and DnaQ‐H domains from His6‐tag (A, D, F) and StrepII‐tag (B, E, G) affinity columns was used to probe Cas1 and Cas2 interactions. SDS–PAGE fractions of elution peak are shown in the inset.
  1. S‐Cas1 and full‐length Cas2‐DnaQ‐H isolation on His6‐tag affinity column.
  2. S‐Cas1 and full‐length Cas2‐DnaQ‐H isolation on StrepII affinity column.
  3. S‐Cas1 and Cas2‐DnaQ‐H proteins elute as a single peak from size‐exclusion column.
  4. S‐Cas1 and Cas2‐H domain isolation on His6‐tag affinity column.
  5. S‐Cas1 and Cas2‐H isolation on StrepII affinity column.
  6. When S‐Cas1 was co‐expressed with DnaQ‐H, only DnaQ‐H was isolated on His6‐tag affinity column.
  7. When S‐Cas1 was co‐expressed with DnaQ‐H, only S‐Cas1 was isolated on StrepII affinity column.