BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1024

Peptides:

Sequences

Affinity Values

QRGRKA(1)
HYGRSG(1)
ERARGA(1)
ALRRGD(1)
DYRGRM(1)
ERLRKA(1)
FGRHAA(1)
FLPRTA(1)
FRGRAA(1)
HRMRMG(1)
IMRRGK(1)
ITYGRR(1)
KFTRSG(1)
LIPRRA(1)
MTRKRM(1)
NFARMG(1)
NHLRKA(1)
NVGRMG(1)
NVSRRG(1)
PISRRA(1)
PVGRMG(1)
RLLRSV(1)
SFGRRH(1)
SLRGRS(1)
TVLRRA(1)
VARRVK(1)
VIARSN(1)
VNTKSG(1)
VRARGA(1)
VRRGRS(1)
VRRRGA(1)
TRVRAK(1)

[430]
[322]
[52]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]
[NT]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 9195973

Target Name:

Tissue-type plasminogen activator

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

X6 fAFF1-tether C phage display library

Affinity Measurement Method:

Dot blot,Binding assay

Affinity Measurement Description:

As the results of the dot blot assay, phage 7 containing the hexamer FRGRAA was a t-PA-selective substrate. The dot blot assay can rapidly provide information regarding both the activity and specificity of individual substrate phage clones. Based on the results of these assays, three peptide substrates (ERARGA, HYGRSG and QRGRKA) were synthesized and characterized to provide a quantitative analysis of the properties of putative t-PA-selective substrates. These peptides were cleaved 180-1500-fold times more efficiently by t-PA than a control peptide (kcat/Km, 0.29 1/ms) containing the physiological cleavage site present in plasminogen. Kcat/Km (1/ms) were shown. NT denotes not tested.

Brief Description:

The initial random hexapeptide library fAFF-TC-LIB was subjected to three rounds of high stringency screening with t-PA to prepare an intermediate library containing phage whose randomized hexamer sequences were digested efficiently by t-PA. The intermediate library was then amplified and screened, at low stringency, with u-PA. Following digestion of the intermediate library with u-PA, mAb E-7 and immobilized protein A were added to the mixture, and the resulting ternary complexes were pelleted by centrifugation. The precipitated ternary complexes were retained and the supernatant were discarded, i.e. phage digested by u-PA were substracted.

BiopanningDataSet Comments:

Curator:

Curation Date:

0000-00-00

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