@article {46065,
	title = {Structural insights into the unique inhibitory mechanism of Kunitz type trypsin inhibitor from Cicer arietinum L. Vol. 37},
	journal = {Journal of Biomolecular Structure \& Dynamics},
	volume = {37},
	year = {2019},
	month = {JUL},
	pages = {2669-2677},
	type = {Article},
	abstract = {<p>Kunitz-type trypsin inhibitors bind to the active pocket of trypsin causing its inhibition. Plant Kunitz-type inhibitors are thought to be important in defense, especially against insect pests. From sequence analysis of various Kunitz-type inhibitors from plants, we identified CaTI2 from chickpea as a unique variant lacking the functionally important arginine residue corresponding to the soybean trypsin inhibitor (STI) and having a distinct and unique inhibitory loop organization. To further explore the implications of these sequence variations, we obtained the crystal structure of recombinant CaTI2 at 2.8 angstrom resolution. It is evident from the structure that the variations in the inhibitory loop facilitates non-substrate like binding of CaTI2 to trypsin, while the canonical inhibitor STI binds to trypsin in substrate like manner. Our results establish the unique mechanism of trypsin inhibition by CaTI2, which warrant further research into its substrate spectrum.</p>
},
	keywords = {Chickpea, crystal structure, inhibitory loop, Kunitz trypsin inhibitor, Trypsin},
	issn = {0739-1102},
	doi = {10.1080/07391102.2018.1494633},
	author = {Bendre, Ameya D. and Suresh, C. G. and Shanmugam, Dhanasekaran and Ramasamy, Sureshkumar}
}
@article {46785,
	title = {Functional stability and structural transitions of a kunitz trypsin inhibitor from chickpea (CaTI2)},
	journal = {Protein Journal},
	volume = {39},
	year = {2020},
	month = {AUG},
	pages = {350-357},
	type = {Article},
	abstract = {<p>Enzymes are important tools for various applications. We have studied structural transitions and functional stability of a Kunitz trypsin inhibitor from Chickpea (CaTI2), a potent insect gut-protease inhibitor, under different stress conditions like non-neutral pH, elevated temperature and co-solvent concentrations. CaTI2 was cloned and expressed in an eukaryotic systemP. pastorisand was investigated for conformational transitions using circular dichroism spectroscopy, differential scanning fluorimetry and activity assay. Native CaTI2 has a sheet dominant structure with 40\% beta sheets and possess a single tryptophan residue situated in the hydrophobic core of the enzyme. The recombinant inhibitor maintained its maximum activity under alkaline pH with its secondary structure intact between pH 6-10. CaTI2 was observed to be thermally stable up to 55 degrees C with aT(m) of 61.3 degrees C above which the protein unfolds. On treating with chemical denaturant (urea), the CaTI2 lost its inhibitory potential and native conformation beyond 2 M urea concentration. Moreover, the protein unfolded at lower temperatures as the concentration of denaturant increased, suggesting more complex structural changes. Further, the stability of the inhibitor was found to be directly proportional to the solvent polarity. The data, herein offers significant information of inhibitor stability and activity which could be exploited for its further development into an effective pesticide. [GRAPHICS] .</p>
},
	keywords = {Chickpea, Circular dichroism spectroscopy, Differential scanning fluorimetry, Functional stability, Kunitz trypsin inhibitor},
	issn = {1572-3887},
	doi = {10.1007/s10930-020-09911-2, Early Access Date = JUL 2020},
	author = {Bendre, Ameya D. and Shukla, Ekta and Ramasamy, Sureshkumar}
}
