English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
 
 
DownloadE-Mail
  Crystal Structure of Crataeva tapia Bark Protein (CrataBL) and Its Effect in Human Prostate Cancer Cell Lines

Ferreira, R. d. S., Zhou, D., Ferreira, J. G., Cabral Silva, M. C., Silva-Lucca, R. A., Mentele, R., et al. (2013). Crystal Structure of Crataeva tapia Bark Protein (CrataBL) and Its Effect in Human Prostate Cancer Cell Lines. PLOS ONE, 8(6): e64426. doi:10.1371/journal.pone.0064426.

Item is

Files

show Files
hide Files
:
pone.0064426.pdf (Any fulltext), 3MB
Name:
pone.0064426.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
open access article
License:
-

Locators

show

Creators

show
hide
 Creators:
Ferreira, Rodrigo da Silva1, Author
Zhou, Dongwen1, Author
Ferreira, Joana Gasperazzo1, Author
Cabral Silva, Mariana Cristina1, Author
Silva-Lucca, Rosemeire Aparecida1, Author
Mentele, Reinhard2, Author              
Paredes-Gamero, Edgar Julian1, Author
Bertolin, Thiago Carlos1, Author
dos Santos Correia, Maria Tereza1, Author
Guedes Paiva, Patricia Maria1, Author
Gustchina, Alla1, Author
Wlodawer, Alexander1, Author
Vilela Oliva, Maria Luiza1, Author
Affiliations:
1external, ou_persistent22              
2Lottspeich, Friedrich / Protein Analysis, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565158              

Content

show
hide
Free keywords: TRYPSIN-INHIBITOR; SERINE RESIDUE; REACTIVE SITE; BINDING; LECTIN; APOPTOSIS; MECHANISM; SEEDS; DU145; GLYCOSAMINOGLYCANS
 Abstract: A protein isolated from the bark of Crataeva tapia (CrataBL) is both a Kunitz-type plant protease inhibitor and a lectin. We have determined the amino acid sequence and three-dimensional structure of CrataBL, as well as characterized its selected biochemical and biological properties. We found two different isoforms of CrataBL isolated from the original source, differing in positions 31 (Pro/Leu); 92 (Ser/Leu); 93 (Ile/Thr); 95 (Arg/Gly) and 97 (Leu/Ser). CrataBL showed relatively weak inhibitory activity against trypsin (K-iapp = 43 mu M) and was more potent against Factor Xa (K-iapp = 8.6 mu M), but was not active against a number of other proteases. We have confirmed that CrataBL contains two glycosylation sites and forms a dimer at high concentration. The high-resolution crystal structures of two different crystal forms of isoform II verified the beta-trefoil fold of CrataBL and have shown the presence of dimers consisting of two almost identical molecules making extensive contacts (similar to 645 angstrom(2)). The structure differs from those of the most closely related proteins by the lack of the N-terminal beta-hairpin. In experiments aimed at investigating the biological properties of CrataBL, we have shown that addition of 40 mM of the protein for 48 h caused maximum growth inhibition in MTT assay (47% of DU145 cells and 43% of PC3 cells). The apoptosis of DU145 and PC3 cell lines was confirmed by flow cytometry using Annexin V/FITC and propidium iodide staining. Treatment with CrataBL resulted in the release of mitochondrial cytochrome c and in the activation of caspase-3 in DU145 and PC3 cells.

Details

show
hide
Language(s): eng - English
 Dates: 2013
 Publication Status: Published online
 Pages: 14
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: PLOS ONE
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA : PUBLIC LIBRARY SCIENCE
Pages: - Volume / Issue: 8 (6) Sequence Number: e64426 Start / End Page: - Identifier: ISSN: 1932-6203