A Bowman-Birk protease inhibitor purified, cloned, sequenced and characterized from the seeds of Maclura pomifera (Raf.) Schneid (original) (raw)
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Recent Updates on Pharmaceutical Potential of Plant Protease Inhibitors
TJPRC, 2013
Plants are rich source of chemicals that are helpful for human in many ways. The pharmacological properties of several different classes of secondary metabolic compounds isolated from different groups of plants are well known. But there are some other compounds that are different from so called secondary metabolic compounds, are also beneficial from pharmacological point of view. One of such compound is small molecular weight proteins that act as inhibitors of proteases. Proteases are essential in survival and maintenance of living organisms. But when the amount increases much higher than the optimum value, they can damage the living cells and tissues. Thus, the involvement of protease inhibitors is essentially required to regulate the amount of proteases within the living cells and tissues. In plants proteases inhibitors are helpful in protecting them from the deleterious effect of insect gut proteases and thus provide defense against insect pests. Several groups of plant protease inhibitors along with their sequences are reported. Many inhibitors are being over expressed in transgenic plants for better insecticidal property. Besides, plant protease inhibitors have enormous pharmaceutical value which is still less explored. The current knowledge of plant protease inhibitors and their pharmaceutical applications have been reviewed.
Analysis of the amino acid sequences of plant Bowman-Birk inhibitors
Journal of Molecular Evolution, 1996
Plant seeds contain a large number of protease inhibitors of animal, fungal, and bacterial origin. One of the well-studied families of these inhibitors is the Bowman-Birk family(BBI). The BBIs from dicotyledonous seeds are 8K, double-headed proteins. In contrast, the 8K inhibitors from monocotyledonous seeds are single headed. Monocots also have a 16K, double-headed inhibitor. We have determined the primary structure of a Bowman-Birk inhibitor from a dicot, horsegram, by sequential edman analysis of the intact protein and peptides derived from enzymatic and chemical cleavage. The 76-residue-long inhibitor is very similar to that ofMacrotyloma axillare. An analysis of this inhibitor along with 26 other Bowman-Birk inhibitor domains (MW 8K) available in the SWISSPROT databank revealed that the proteins from monocots and dicots belong to related but distinct families. Inhibitors from monocots show larger variation in sequence. Sequence comparison shows that a crucial disulphide which connects the amino and carboxy termini of the active site loop is lost in monocots. The loss of a reactive site in monocots seems to be correlated to this. However, it appears that this disulphide is not absolutely essential for retention of inhibitory function. Our analysis suggests that gene duplication leading to a 16K inhibitor in monocots has occurred, probably after the divergence of monocots and dicots, and also after the loss of second reactive site in monocots.
In Vitro Screening of Seed Extracts of Medicinal Plants for Protease Inhibitory Activity
Cihan University-Erbil Scientific Journal
Protease inhibitors (PIs) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. In humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. The present investigation reports PIs activity data from 30 medicinal plants. The screening for PIs activity was done by dot blot assay using X-ray film coated with gelatin. Among screened seed extracts, Albizia lebbeck, Raphanus sativus, Mucuna pruriens, Achyranthes aspera, and Coffea arabica showed high inhibitory activities with trypsin protease. Most of seed extracts exhibited moderate activity, whereas Ocimum sanctum showed moderate to low activity against trypsin. The presence of varied protein content is reported from all seed extracts with highest in A. lebbeck (50.0 ± 3.4 mg/ml). The data produ...
Cysteine protease inhibitors (CPIs) have been known to be present in a variety of seeds of plants, and have been intensively studied as useful tools for potential utilization in pharmacology. This study reports the isolation of CPI from Tetracarpidium conophorum by 65% ammonium sulphate saturation, followed by ion exchange chromatography; further purification was by gel filtration chromatography. The molecular weight of the partially purified protein inhibitor was analyzed by SDS-PAGE to be approximately 20 kDa. The inhibitor had an optimum pH and temperature of 8.0 and 40°C, respectively. The inhibitor competitively inhibited papain with the same Vmax = 71.1710 3 µmol/min, Km = 166 µM, and Ki = 53.63 µM. Divalent metal ions such as, Mg 2+ , Pb 2+ , Mn 2+ , Cu 2+ , Co 2+ , and Zn 2+ had significant effect on inhibitory activity of CPI at concentration as low as 1 mM. Cysteine protease inhibitor of T. conophorum investigated in this study could serve as a template in biotechnology o...
Frontiers in Plant Science, 2021
A Bowman-Birk protease, i.e., Mucuna pruriens trypsin inhibitor (MPTI), was purified from the seeds by 55.702-fold and revealed a single trypsin inhibitor on a zymogram with a specific activity of 202.31 TIU/mg of protein. On sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) under non-reducing conditions, the protease trypsin inhibitor fraction [i.e., trypsin inhibitor non-reducing (TINR)] exhibited molecular weights of 74 and 37 kDa, and under reducing conditions [i.e., trypsin inhibitor reducing (TIR)], 37 and 18 kDa. TINR-37 revealed protease inhibitor activity on native PAGE and 37 and 18 kDa protein bands on SDS–PAGE. TINR-74 showed peaks corresponding to 18.695, 37.39, 56.085, and 74.78 kDa on ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization/quadrupole time-of-flight-mass spectrometry (ESI/QTOF-MS). Similarly, TINR-37 displayed 18.695 and 37.39 kDa peaks. Furthermore, TIR-37 and TIR-18 exhibited peaks corresponding to 37.39...