A prolyl oligopeptidase inhibitor, Z-Pro-Prolinal, inhibits glyceraldehyde-3-phosphate dehydrogenase translocation and production of reactive oxygen species in CV1-P cells exposed to 6-hydroxydopamine.

Abstract

We studied the ability of prolyl oligopeptidase (POP) inhibitors, Z-Pro-Prolinal and JTP-4819, to prevent translocation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and formation of reactive oxygen species (ROS), in 6-hydroxydopamine (6-OHDA) and cytosine arabinoside (Ara-C) treated monkey fibroblast (CV1-P) and human neuroblastoma (SH-SY5Y) cells. The cells were pretreated with POP inhibitors (30 min) before addition of toxicants. GAPDH was analyzed by Western hybridization, ROS by fluorescent 2'7'-dichlorodihydro-fluorescein diacetate, and viability by the MTT method. Both toxicants induced GAPDH translocation to the particulate fraction (mitochondria and nuclei). Z-Pro-Prolinal was able to inhibit the translocation in 6-OHDA-exposed CV1-P cells. In SH-SY5Y cells and in JTP-4819 pretreated cells, no prevention of translocation was seen. However, the intensity of GAPDH in cytosolic fraction increased. Both inhibitors blocked 6-OHDA-induced ROS-production to the control level in CV1-P but, not in SH-SY5Y cells, without affecting their viability. In conclusion, POP inhibitors are able to prevent certain cell stress related factors such as ROS production or GAPDH translocation.

• Authors: Katja A Puttonen, Sárka Lehtonen, Atso Raasmaja, Pekka T Männistö
• Journal: Toxicology in vitro : an international journal published in association with BIBRA (Toxicol In Vitro) Vol. 20 Issue 8 Pg. 1446-54 (Dec 2006) ISSN: 0887-2333 [Print] England
• PMID: 16942854 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Actins
• Dipeptides
• Protease Inhibitors
• Proto-Oncogene Proteins c-bcl-2
• Pyrrolidines
• Reactive Oxygen Species
• Tetrazolium Salts
• Thiazoles
• Tumor Suppressor Protein p53
• Cytarabine
• N-benzyloxycarbonylprolylprolinal
• Oxidopamine
• JTP 4819
• Glyceraldehyde-3-Phosphate Dehydrogenases
• Serine Endopeptidases
• PREPL protein, human
• Prolyl Oligopeptidases
• thiazolyl blue

Topics

• Actins (biosynthesis, genetics)
• Blotting, Western
• Cell Line
• Cell Survival (drug effects)
• Cytarabine (antagonists & inhibitors, toxicity)
• Dipeptides (pharmacology)
• Glyceraldehyde-3-Phosphate Dehydrogenases (metabolism)
• Humans
• Oxidopamine (antagonists & inhibitors, toxicity)
• Prolyl Oligopeptidases
• Protease Inhibitors (pharmacology)
• Protein Transport (drug effects)
• Proto-Oncogene Proteins c-bcl-2 (metabolism)
• Pyrrolidines (pharmacology)
• Reactive Oxygen Species (metabolism)
• Serine Endopeptidases (metabolism)
• Tetrazolium Salts
• Thiazoles
• Tumor Suppressor Protein p53 (metabolism)