Molecular cloning of Kunitz-type proteinase inhibitor group B genes from potato

Speranskaya, A.S.; Krinitsina, A.A.; Poltronieri, Palmiro; Fasano, Pasqua; Santino, Angelo; Шевелев, А. Б.; Valueva, T. A.

doi:10.1007/s10541-005-0114-3

Cited by 7 publications

(1 citation statement)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further, the KTI family showed unique sequence variation, including many nonsynonymous substitutions and loops, which appear to translate into functional diversity in the plants. [51]. 3 novel genes such as PKPI-B1, PKPI-B2, and PKPI-B10 represented by five, two, and seven clones, respectively.…”

Section: Purification and Characterization Of Solanum Pimentioning

confidence: 99%

Solanum Protease Inhibitors and Their Therapeutic Potentialities: A Review

Murugan

2016

Int J Pharm Pharm Sci

View full text Add to dashboard Cite

Protease inhibitors (PIs) are diverse group of proteins with low molecular weight that are ubiquitous in all life forms. PIs are reducers of the physiological activity of proteases and fascinated the attention of biotechnological researchers. In the evolutionary course, plants have developed diverse adaptive mechanisms of defence against various unfavourable conditions including that of predators and pathogens. Phylogenetic relationships among diverse PI families like serpin, Bowman-Birk, cereal trypsin/α-amylase inhibitor, proteinase inhibitor I, proteinase inhibitor II and cystatin have been evaluated. PIs evolution seems to occur through multiple interacting mechanisms and not commonly seen with other coevolving molecules. Interaction of PIs produced by host organisms and the invasive/dietary protease of pathogens or parasites or predators, leads to a phylogenetic 'arms race' of rapid structural modulation in both interacting proteins. Further, the high rate of retention of gene duplication and inhibitory domain multiplication results the PI as potential model system to trace the basic evolutionary process of functional diversification. The mode of action of PI is either via inactivating the hydrolytic enzymes or depolarization of cell membrane of the pathogens thereby inhibiting its growth and invasion. Generally, PIs possess significant number of disulfide bonds due to cysteine residues that provide them resistance to heat, extremes of pH, and proteolysis. However, PIs have been extracted and purified only from few monocots and dicots plants. Currently, PI genes were used for developing insect-resistant transgenic plants for crop improvements. Classification of PIs over the last several years has been based on structural-functional relationships. This review bridges the gap between the folkloric uses of Solanum PIs, their diversity and biological potentialities.

show abstract

Section: Purification and Characterization Of Solanum Pimentioning

confidence: 99%

Solanum Protease Inhibitors and Their Therapeutic Potentialities: A Review

Murugan

2016

Int J Pharm Pharm Sci

View full text Add to dashboard Cite

show abstract

Kunitz‐type protease inhibitors group B from Solanum palustre

Speransky

Cimaglia

Krinitsina

et al. 2007

Biotechnology Journal

View full text Add to dashboard Cite

Five Kunitz protease inhibitor group B genes were isolated from the genome of the diploid non-tuber-forming potato species Solanum palustre. Three of five new genes share 99% identity to the published KPI-B genes from various cultivated potato accessions, while others exhibit 96% identity. Spls-KPI-B2 and Spls-KPI-B4 proteins contain unique substitutions of the most conserved residues usually involved to trypsin and chymotrypsin-specific binding sites of Kunitz-type protease inhibitor (KPI)-B, respectively. To test the inhibition of trypsin and chymotrypsin by Spls-KPI proteins, five of them were produced in E. coli purified using a Ni-sepharose resin and ion-exchange chromatography. All recombinant Spls-KPI-B inhibited trypsin; K(i) values ranged from 84.8 (Spls-KPI-B4), 345.5 (Spls-KPI-B1), and 1310.6 nM (Spls-KPI-B2) to 3883.5 (Spls-KPI-B5) and 8370 nM (Spls-KPI-B3). In addition, Spls-KPI-B1 and Spls-KPI-B4 inhibited chymotrypsin. These data suggest that regardless of substitutions of key active-center residues both Spls-KPI-B4 and Spls-KPI-B1 are functional trypsin-chymotrypsin inhibitors.

show abstract