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(a) Schematic of the genetic engineering strategy for the generation of the ChuA reporter strain used in this study. (b) The growth phenotype of the strain described in (a), grown on LB agar (top, representative images shown) in the presence of iron-limited LB agar supplemented with either 5 µM αβHb, αγHb, hemin, myoglobin, <t>neuroglobin,</t> cytoglobin or human ferredoxin 1. (below) EC 50 values of E. coli ΔTBDT:ChuOP cultured in LB liquid medium in the presence of 200 µM 2,2’-bipyridine, supplemented with serially diluted αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or ferredoxin 1.
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(a) Schematic of the genetic engineering strategy for the generation of the ChuA reporter strain used in this study. (b) The growth phenotype of the strain described in (a), grown on LB agar (top, representative images shown) in the presence of iron-limited LB agar supplemented with either 5 µM αβHb, αγHb, hemin, myoglobin, <t>neuroglobin,</t> cytoglobin or human ferredoxin 1. (below) EC 50 values of E. coli ΔTBDT:ChuOP cultured in LB liquid medium in the presence of 200 µM 2,2’-bipyridine, supplemented with serially diluted αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or ferredoxin 1.
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(a) Schematic of the genetic engineering strategy for the generation of the ChuA reporter strain used in this study. (b) The growth phenotype of the strain described in (a), grown on LB agar (top, representative images shown) in the presence of iron-limited LB agar supplemented with either 5 µM αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or human ferredoxin 1. (below) EC 50 values of E. coli ΔTBDT:ChuOP cultured in LB liquid medium in the presence of 200 µM 2,2’-bipyridine, supplemented with serially diluted αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or ferredoxin 1.

Journal: bioRxiv

Article Title: Inhibiting heme-piracy by pathogenic Escherichia coli using de novo -designed proteins

doi: 10.1101/2024.12.05.626953

Figure Lengend Snippet: (a) Schematic of the genetic engineering strategy for the generation of the ChuA reporter strain used in this study. (b) The growth phenotype of the strain described in (a), grown on LB agar (top, representative images shown) in the presence of iron-limited LB agar supplemented with either 5 µM αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or human ferredoxin 1. (below) EC 50 values of E. coli ΔTBDT:ChuOP cultured in LB liquid medium in the presence of 200 µM 2,2’-bipyridine, supplemented with serially diluted αβHb, αγHb, hemin, myoglobin, neuroglobin, cytoglobin or ferredoxin 1.

Article Snippet: Sequences for the expression of recombinant human αβ and αγ hemoglobin, myoglobin, neuroglobin, cytoglobin and ferredoxin 1 were ordered as gene fragments from Twist Biosciences and cloned into either pETDuet-1 (for hemoglobin), pET29a or pET22b, carrying an N-terminal 6x His tag.

Techniques: Cell Culture