BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 44

Peptides:

Sequences
TTSPLSQGSSYI(9)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 16369013

Target Name:

Listeria monocytogenes

Template Name:

Cation-independent mannose-6-phosphate receptor

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Panning was performed using Dynex Immunolon microtiter wells coated with L. monocytogenes cocktail (15 serotypes or strains, i.e. 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, 4a, EGD and three EGD Mutant strains). Nonspecific phages were removed by absorbing the phage library sequentially on uncoated wells and wells coated with BSA. Subtractive selection was carried out by the sequential application of L. innocua cocktail and L. ivanovii cocktail to wells with washing between steps. Low-affinity L. monocytogenes-specific phage peptides were removed by rapid exposure and removal of L. monocytogenes cocktail. The wells were washed before incubation with L. monocytogenes cocktail (30 min) to elute specific high-affinity bound phages.

BiopanningDataSet Comments:

TTSPLSQGSSYI is quite similar to the segment PLAQSGGSSYI in the human cation-independent mannose 6-phosphate receptor (M6PR), which is within the mannose 6-phosphate binding site of the M6PR. Binding of the labeled synthetic PLAQSGGSSYI peptide by Listeria spp. was confirmed by fluorescence spectrophotometry and FACS. Further evidence for binding of the receptor by L. monocytogenes and L. innocua was provided by affinity purification. M6PR is thus indentified as a novel receptor for binding and invasion of Listeria species.

Curator:

Curation Date:

0000-00-00

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