TY - JOUR
T1 - Development of novel peptide inhibitor of Lipoxygenase based on biochemical and BIAcore evidences
AU - Somvanshi, Rishi Kumar
AU - Singh, Abhay Kumar
AU - Saxena, Meera
AU - Mishra, Biswajit
AU - Dey, Sharmistha
PY - 2008/11
Y1 - 2008/11
N2 - Lipoxygenase (LOX) are enzymes implicated in a broad range of inflammatory diseases, cancer, asthma and atherosclerosis. These diverse biological properties lead to the interesting target for the inhibition of this metabolic pathway of LOX. The drugs available in the market against LOX reported to have various side effects. To develop potent and selective therapeutic agents against LOX, it is essential to have the knowledge of its active site. Due to the lack of structural data of human LOX, researchers are using soybean LOX (sLOX) because of their availability and similarities in the active site structure. Based on the crystal structure of sLOX-3 and its complex with known inhibitors, we have designed a tripeptide, FWY which strongly inhibits sLOX-3 activity. The inhibition by peptide has been tested with purified sLOX-3 and with LOX present in blood serum of breast cancer patients in the presence of substrate linoleic acid and arachidonic acid respectively. The dissociation constant (KD) of the peptide with sLOX-3 as determined by Surface Plasmon Resonance (SPR) was 3.59 × 10- 9 M. The kinetic constant (Ki) and IC50, as determined biochemical methods were 7.41 × 10- 8 M and 0.15 × 10- 6 M respectively.
AB - Lipoxygenase (LOX) are enzymes implicated in a broad range of inflammatory diseases, cancer, asthma and atherosclerosis. These diverse biological properties lead to the interesting target for the inhibition of this metabolic pathway of LOX. The drugs available in the market against LOX reported to have various side effects. To develop potent and selective therapeutic agents against LOX, it is essential to have the knowledge of its active site. Due to the lack of structural data of human LOX, researchers are using soybean LOX (sLOX) because of their availability and similarities in the active site structure. Based on the crystal structure of sLOX-3 and its complex with known inhibitors, we have designed a tripeptide, FWY which strongly inhibits sLOX-3 activity. The inhibition by peptide has been tested with purified sLOX-3 and with LOX present in blood serum of breast cancer patients in the presence of substrate linoleic acid and arachidonic acid respectively. The dissociation constant (KD) of the peptide with sLOX-3 as determined by Surface Plasmon Resonance (SPR) was 3.59 × 10- 9 M. The kinetic constant (Ki) and IC50, as determined biochemical methods were 7.41 × 10- 8 M and 0.15 × 10- 6 M respectively.
KW - Enzyme kinetics
KW - Leukotrienes
KW - Lipoxygenases
KW - Peptide synthesis
KW - Surface plasmon resonance
UR - http://www.scopus.com/inward/record.url?scp=54049149323&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2008.07.004
DO - 10.1016/j.bbapap.2008.07.004
M3 - Article
C2 - 18691678
AN - SCOPUS:54049149323
SN - 1570-9639
VL - 1784
SP - 1812
EP - 1817
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
IS - 11
ER -