schliessen

Filtern

 

Bibliotheken

Comparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors

Aim and Objective: Human full-length cyclic nucleotide phosphodiesterase isozyme 4B2 (hPDE4B2) as the target for screening and characterizing inhibitors suffers from low activity yield and the coexistence of two conformational states bearing different affinities for (R)-rolipram. Hence, the 152~528... Full description

Journal Title: Combinatorial Chemistry & High Throughput Screening 2019, Vol.22(1), p.49-58
Main Author: Zhang, Xiang
Other Authors: He, Shu , Hu, Xiaolei , Wu, Jing , Li, Xinpeng , Liao, Fei , Yang, Xiaolan
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 1386-2073 ; E-ISSN: 1875-5402 ; DOI: 10.2174/1386207322666190306142810
Link: http://www.eurekaselect.com/openurl/content.php?genre=article&issn=13862073&volume=22&issue=1&spage=49
Zum Text:
SendSend as email Add to Book BagAdd to Book Bag
Staff View
recordid: bentham10.2174/1386207322666190306142810
title: Comparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors
format: Article
creator:
  • Zhang, Xiang
  • He, Shu
  • Hu, Xiaolei
  • Wu, Jing
  • Li, Xinpeng
  • Liao, Fei
  • Yang, Xiaolan
subjects:
  • Human Cyclic Nucleotide Phosphodiesterase 4b Truncate Inhibitor Rolipram Sumo Tag Inflammatory.
ispartof: Combinatorial Chemistry & High Throughput Screening, 2019, Vol.22(1), p.49-58
description: Aim and Objective: Human full-length cyclic nucleotide phosphodiesterase isozyme 4B2 (hPDE4B2) as the target for screening and characterizing inhibitors suffers from low activity yield and the coexistence of two conformational states bearing different affinities for (R)-rolipram. Hence, the 152~528 truncate of hPDE4B2 existing only in the low-affinity conformation state for (R)-rolipram was compared against the full-length hPDE4B2 to characterize inhibitors.
Materials and Methods: With 6His-SUMO tag at the N-terminus, both the full-length hPDE 4B2 (SF-hPDE4B2) and the 152~528 truncate (ST-hPDE4B2) were expressed in Escherichia coli cells, purified through Ni-NTA column and compared for the characterization of inhibitors. The inhibition constants (Ki) of some synthesized rolipram analogues against both targets were determined with 96-well microplate through the coupled action of monophosphatase on AMP and spectrophotometric assay of phosphate with malachite green.
Results: After affinity purification with Ni2+-NTA column, ST-hPDE4B2 showed about 30-fold higher specific activity and 100-fold higher activity yield than SF-hPDE4B2; Ki of (R)-rolipram on ST-hPDE4B2 was consistent with that on the low-affinity state of the untagged full-length hPDE4B2 expressed in insect cells. Of some representative rolipram analogues as inhibitors, a dual-logarithm model quantitatively described their monotonic association, and Ki from 0.010 mM to 8.5 mM against SF-hPDE4B2 was predicted from Ki against ST-hPDE4B2, supporting the discovery of consistent hits by the use of both targets with a pair of properly-set cutoffs.
Conclusion: ST-hPDE4B2 with much higher activity yield may be a favorable alternative target to characterize/screen rolipram analogues as hPDE4B inhibitors in high-throughput mode.
language: eng
source:
identifier: ISSN: 1386-2073 ; E-ISSN: 1875-5402 ; DOI: 10.2174/1386207322666190306142810
fulltext: fulltext
issn:
  • 1386-2073
  • 13862073
  • 1875-5402
  • 18755402
url: Link


@attributes
ID1047950005
RANK0.06999999
NO1
SEARCH_ENGINEprimo_central_multiple_fe
SEARCH_ENGINE_TYPEPrimo Central Search Engine
LOCALfalse
PrimoNMBib
record
control
sourcerecordid10.2174/1386207322666190306142810
sourceidbentham
recordidTN_bentham10.2174/1386207322666190306142810
sourceformatXML
sourcesystemPC
pqid2188985507
display
typearticle
titleComparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors
creatorZhang, Xiang ; He, Shu ; Hu, Xiaolei ; Wu, Jing ; Li, Xinpeng ; Liao, Fei ; Yang, Xiaolan
ispartofCombinatorial Chemistry & High Throughput Screening, 2019, Vol.22(1), p.49-58
identifier
subjectHuman Cyclic Nucleotide Phosphodiesterase 4b Truncate Inhibitor Rolipram Sumo Tag Inflammatory.
description
languageeng
source
version6
lds50peer_reviewed
links
openurl$$Topenurl_article
backlink$$Uhttp://www.eurekaselect.com/openurl/content.php?genre=article&issn=13862073&volume=22&issue=1&spage=49$$EView_record_at_Bentham_Science
openurlfulltext$$Topenurlfull_article
search
creatorcontrib
0Zhang, Xiang
1He, Shu
2Hu, Xiaolei
3Wu, Jing
4Li, Xinpeng
5Liao, Fei
6Yang, Xiaolan
titleComparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors
description
subject
general
010.2174/1386207322666190306142810
1Bentham Science Publishers Ltd.
2English
3Bentham Science - Journals
sourceidbentham
recordidbentham10.2174/1386207322666190306142810
issn
01386-2073
113862073
21875-5402
318755402
rsrctypearticle
creationdate2019
addtitleCombinatorial Chemistry & High Throughput Screening
searchscopebentham
scopebentham
lsr30VSR-Enriched:[pqid]
sort
titleComparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors
authorZhang, Xiang ; He, Shu ; Hu, Xiaolei ; Wu, Jing ; Li, Xinpeng ; Liao, Fei ; Yang, Xiaolan
creationdate20190000
facets
frbrgroupid2572723724577078643
frbrtype5
newrecords20190521
languageeng
creationdate2019
topic
collectionBentham Science - Journals
prefilterarticles
rsrctypearticles
creatorcontrib
0Zhang, Xiang
1He, Shu
2Hu, Xiaolei
3Wu, Jing
4Li, Xinpeng
5Liao, Fei
6Yang, Xiaolan
jtitleCombinatorial Chemistry & High Throughput Screening
toplevelpeer_reviewed
delivery
delcategoryRemote Search Resource
fulltextfulltext
addata
aulast
0Zhang
1He
2Hu
3Wu
4Li
5Liao
6Yang
aufirst
0Xiang
1Shu
2Xiaolei
3Jing
4Xinpeng
5Fei
6Xiaolan
au
0Zhang, Xiang
1He, Shu
2Hu, Xiaolei
3Wu, Jing
4Li, Xinpeng
5Liao, Fei
6Yang, Xiaolan
atitleComparison of the Full-Length and 152~528 Truncate of Human Cyclic Nucleotide Phosphodiesterase 4B2 for the Characterization of Inhibitors
jtitleCombinatorial Chemistry & High Throughput Screening
date2019
risdate2019
volume22
issue1
spage49
epage58
pages49-58
issn1386-2073
eissn1875-5402
formatjournal
genrearticle
ristypeJOUR
abstract
pubBentham Science Publishers Ltd.
doi10.2174/1386207322666190306142810