BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1948

Peptides:

Sequences
TLHLHHL(15)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 22372912

Target Name:

Transcriptional regulatory protein devR (dosR)

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-7 phage display library (X7)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

In the first round, the phage library was incubated with (His)6-DevR immobilized on Ni2+ NTA (nitrilotriacetic acid) magnetic agarose beads. The loosely bound phages were discarded by elution with DevR D54N mutant protein (100μg/mL concentration), which differs from the wild-type protein at the phosphorylation site. A second elution was performed with buffer containing 250 mM imidazole that released the His6-tagged protein along with the tightly bound phages. Three rounds of panning were performed using the twin elutions approach, and each time the phages in the imidazole elution were amplified and used as an input for the next round of panning. The fourth and fifth rounds were performed on plate to eliminate bead binders. The loosely bound phages were\r\nfirst eluted with mutant D54N DevR protein and then with 0.2 M glycine, pH 2.2. In the fifth (final) round of panning, a penultimate elution using phosphorylated DevS was carried out prior to final elution of the bound phages using 0.2 M glycine.

BiopanningDataSet Comments:

DevRS1 (TLHLHHL) inhibited DevR-regulated transcription and survival of nonreplicating tubercle bacilli in a hypoxia model of dormancy. DevRS1 peptide-mediated inhibition demonstrates the efficacy of intercepting DevR function to block hypoxic adaptation of M. tb.

Curator:

Curation Date:

0000-00-00

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