PMID- 17156847
OWN - NLM
STAT- MEDLINE
DCOM- 20070516
LR  - 20070312
IS  - 0165-022X (Print)
IS  - 0165-022X (Linking)
VI  - 70
IP  - 3
DP  - 2007 Apr 10
TI  - Construction of a high sensitive Escherichia coli alkaline phosphatase reporter 
      system for screening affinity peptides.
PG  - 435-9
AB  - An enzyme-linker-peptide fusion protein reporter system was constructed for 
      sensitive analysis of affinity of peptide ligands to their receptor. An E. coli 
      alkaline phosphatase (EAP) mutant enzyme with high catalytic activity was 
      selected as the reporter protein. Interaction of affinity peptide and 
      streptavidin was applied as demonstration of the method. Three affinity peptides, 
      strep-tag I (SI), strep-tag II (SII) and streptavidin binding peptide (SBP) were 
      genetically fused to the C-terminal of EAP respectively, with an insertion of a 
      flexible linker peptide in between. The enzyme activity of the EAP fusions showed 
      no obvious change. After expression and purification, the EAP-affinity peptide 
      fusions were applied to the streptavidin modified surface. Binding of the fusions 
      to the surface through interaction of affinity peptides to streptavidin was 
      indicated by color generated from conversion of the substrate by EAP. The 
      relative affinity and specificity of each affinity peptides to the immobilized 
      streptavidin were then evaluated with high sensitivity and broad detection range. 
      This method may be used for effective high-throughput screening of high affinity 
      peptide from the peptide pool.
FAU - Huang, Xu
AU  - Huang X
AD  - State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of 
      Sciences, Wuhan 430071, China.
FAU - Zhang, Xian-En
AU  - Zhang XE
FAU - Zhou, Ya-Feng
AU  - Zhou YF
FAU - Zhang, Zhi-Ping
AU  - Zhang ZP
FAU - Cass, Anthony E G
AU  - Cass AE
LA  - eng
PT  - Journal Article
DEP - 20061021
PL  - Netherlands
TA  - J Biochem Biophys Methods
JT  - Journal of biochemical and biophysical methods
JID - 7907378
RN  - 0 (Affinity Labels)
RN  - 0 (Ligands)
RN  - 0 (Peptides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - Affinity Labels
MH  - Alkaline Phosphatase/*genetics/metabolism
MH  - Base Sequence
MH  - Escherichia coli/*enzymology/*genetics
MH  - Genes, Reporter
MH  - Genetic Vectors
MH  - Ligands
MH  - *Peptides/metabolism
MH  - Plasmids/genetics
MH  - Protein Binding
MH  - Recombinant Fusion Proteins/genetics/metabolism
EDAT- 2006/12/13 09:00
MHDA- 2007/05/17 09:00
CRDT- 2006/12/13 09:00
PHST- 2006/06/24 00:00 [received]
PHST- 2006/10/10 00:00 [revised]
PHST- 2006/10/10 00:00 [accepted]
PHST- 2006/12/13 09:00 [pubmed]
PHST- 2007/05/17 09:00 [medline]
PHST- 2006/12/13 09:00 [entrez]
AID - S0165-022X(06)00202-8 [pii]
AID - 10.1016/j.jbbm.2006.10.006 [doi]
PST - ppublish
SO  - J Biochem Biophys Methods. 2007 Apr 10;70(3):435-9. doi: 
      10.1016/j.jbbm.2006.10.006. Epub 2006 Oct 21.