Trypsin inhibitors from the garden four o’clock (Mirabilis jalapa) and spinach (Spinacia oleracea) seeds: Isolation, characterization and chemical synthesis
| Autor: | Anna Wilimowska-Pelc, Krzysztof Rolka, Ewa Zabłotna, Katarzyna Pszczola, Adam Lesner, Tadeusz Wilusz, Anna Kwiatkowska, Aleksandra Walewska, Irena Lorenc-Kubis, Małgorzata Sleszyńska, Anna Łęgowska, Mateusz Ługowski, Ewa Zuziak, Jolanta Kowalska |
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| Rok vydání: | 2007 |
| Předmět: |
Spinacia
Molecular Sequence Data Plant Science Horticulture Biochemistry Affinity chromatography Spinacia oleracea Thermolysin medicine Amino Acid Sequence Molecular Biology Plant Proteins Chromatography biology General Medicine Proteinase K biology.organism_classification Trypsin Mirabilis jalapa Seeds biology.protein Spinach Serine Proteinase Inhibitors Mirabilis Trypsin Inhibitors Sequence Alignment medicine.drug |
| Zdroj: | Phytochemistry. 68:1487-1496 |
| ISSN: | 0031-9422 |
| DOI: | 10.1016/j.phytochem.2007.03.012 |
| Popis: | Five serine proteinase inhibitors (Mirabilis jalapa trypsin inhibitors, MJTI I and II and Spinacia oleracea trypsin inhibitors, SOTI I, II, and III) from the garden four-o’clock (M. jalapa) and spinach (S. oleracea) seeds were isolated. The purification procedures included affinity chromatography on immobilized methylchymotrypsin in the presence of 5 M NaCl, ion exchange chromatography and/or preparative electrophoresis, and finally RP–HPLC on a C-18 column. The inhibitors, crosslinked by three disulfide bridges, are built of 35 to 37 amino-acid residues. Their primary structures differ from those of known trypsin inhibitors, but showed significant similarity to the antimicrobial peptides isolated from the seeds of M. jalapa (MJ-AMP1, MJ-AMP2), Mesembryanthemum crystallinum (AMP1), and Phytolacca americana (AMP-2 and PAFP-S) and from the hemolymph of Acrocinus longimanus (Alo-1, 2 and 3). The association equilibrium constants (Ka) with bovine β-trypsin for the inhibitors from M. jalapa (MJTI I and II) and S. oleracea (SOTI I–III) were found to be about 107 M−1. Fully active MJTI I and SOTI I were obtained by solid-phase peptide synthesis. The disulfide bridge pattern in both inhibitors (Cys1–Cys4, Cys2–Cys5 and Cys3–Cys6) was established after their digestion with thermolysin and proteinase K followed by the MALDI-TOF analysis. |
| Databáze: | OpenAIRE |
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