A novel trypsin inhibitor from Peltophorum dubium seeds, with lectin-like properties, triggers rat lymphoma cell apoptosis
2003 (English)In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 411, no 1, 93-104 p.Article in journal (Refereed) Published
A trypsin inhibitor (PDTI) was isolated from Peltophorum dubium seeds by affinity chromatography on a thyroglobulin-agarose or a trypsin-agarose column. In both cases, SDS-PAGE showed two bands of M(r) 20,000 and 22,000, which could not be resolved. Their amino-terminal sequences were identical and similar to that of Kunitz-type soybean trypsin inhibitor (SBTI). Mass spectrometry analysis of tryptic digests of both bands showed 16 coincident peaks, suggesting that they are closely related proteins. The K(i)s for trypsin and chymotrypsin inhibitory activity of PDTI were 1.6 x 10(-7) and 1.3 x 10(-5)M, respectively. Lectin-like activity of PDTI and SBTI, detected by hemagglutination of rabbit erythrocytes, was inhibited by sialic acid-containing compounds. PDTI and SBTI caused apoptosis of Nb2 rat lymphoma cells, demonstrated by decrease of viability, DNA hypodiploidy, DNA fragmentation, and caspase-3-like activity. They had no effect on normal mouse splenocytes or lymphocytes, whereas they caused apoptosis of concanavalin A-stimulated mouse lymphocytes.
Place, publisher, year, edition, pages
2003. Vol. 411, no 1, 93-104 p.
Amino Acid Sequence, Animals, Apoptosis/*drug effects, Cell Survival/drug effects, Chromatography; Affinity, Fabaceae, Lectins, Lymphocytes/drug effects/immunology, Lymphoma/*pathology, Mice, Molecular Sequence Data, Molecular Weight, Peptide Fragments/chemistry, Phytotherapy, Rats, Seeds, Trypsin Inhibitors/chemistry/isolation & purification/*pharmacology, Tumor Cells; Cultured
Natural Sciences Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-10823DOI: 10.1016/S0003-9861(02)00726-9PubMedID: 12590927OAI: oai:DiVA.org:uu-10823DiVA: diva2:38591