Expression and kinetic characterization of barley chymotrypsin inhibitors 1a and 1b

Greagg, Martin A.; Brauer, Arnd B. E.; Leatherbarrow, Robin J.

doi:10.1016/0167-4889(94)90167-8

Cited by 10 publications

(10 citation statements)

References 36 publications

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“…It is supported by solution kinetic data showing that members of both families are inhibited, often with very similar binding constants, by the same protein inhibitors (Read & James, 1986;Bigler et al, 1993;Greagg et al, 1994). Therefore, the subtilisins and chymotrypsins not only have a similar mechanism of catalysis, they also have a similar mechanism of binding to inhibitors, such as eglin c, that obey the standard mechanism of inhibition (Laskowski & Kato, 1980).…”

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confidence: 91%

Interaction of subtilisins with serpins

et al. 1996

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Serpins are well-characterized inhibitors of the chymotrypsin family serine proteinases. We have investigated the interaction of two serpins with members of the subtilisin family, proteinases that possess a similar catalytic mechanism to the chymotrypsins, but a totally different scaffold. We demonstrate that a'proteinase inhibitor inhibits subtilisin Curlsberg and proteinase K, and alantichymotrypsin inhibits proteinase K, but not subtilisin Carlsberg. When inhibition occurs, the rate of formation and stability of the complexes are similar to those formed between serpins and chymotrypsin family members. However, inhibition of subtilisins is characterized by large partition ratios where more than four molecules of each serpin are required to inhibit one subtilisin molecule. The partition ratio is caused by the serpins acting as substrates or inhibitors. The ratio decreases as temperature is elevated in the range 0-45 "C, indicating that the serpins are more efficient inhibitors at high temperature. These aspects of the subtilisin interaction are all observed during inhibition of chymotrypsin family members by serpins, indicating that serpins accomplish inhibition of these two distinct proteinase families by the same mechanism.

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confidence: 91%

Interaction of subtilisins with serpins

et al. 1996

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“…CI-2A and BASI were more potent inhibitors of the SL proteinase than CI-1A. Both CI-1 proteins also inhibited bovine chymotrypsin and bacterial subtilisin more weakly than CI-2A (19). BBBI was a stronger inhibitor than its soybean relative.…”

Section: Discussionmentioning

confidence: 86%

“…The SL proteinase activity was inhibited by the proteins CI-1, CI-2, and BASI. All of these proteins have been previously shown to inhibit proteinases from various nonpathogenic microbes (2,6,9,19,31). Mikola and Suolinna (2) showed that an alkaline proteinase inhibitor, which was later called CI-1 (8), inhibited a proteinase from the phytopathogen Alternaria tenuissima.…”

Section: Discussionmentioning

confidence: 99%

“…These proteins inhibit endogenous seed R-amylases, but their inhibition of microbial amylases has not been detected. CI-1 and CI-2, which belong to the potato inhibitor I family (17,18), inhibit the activities of chymotrypsin, elastase, bacterial subtilisin and certain other microbial proteinases (8,9,19). An inhibitor that is very similar to CI-2A is present in wheat grain endosperm (9,20).…”

Section: Introductionmentioning

confidence: 99%

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Purification and Identification of Barley (Hordeum vulgare L.) Proteins That Inhibit the Alkaline Serine Proteinases of Fusarium culmorum

Pekkarinen

Jones

2003

J. Agric. Food Chem.

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It has been proposed that microbial proteinase inhibitors, which are present in abundance in cereal grains, protect the seed against plant pathogens. So far, however, very little is known about the interactions of those inhibitors with the proteinases of phytopathogenic microbes. The increased alkaline proteinase activities of Fusarium head blight (FHB) diseased wheat and barley grain imply that the Fusarium fungi synthesize those enzymes during the colonization of the kernel. To study which barley proteins can inhibit Fusarium proteinases, and hence, possibly protect the seed from FHB, the proteins of a grain extract have been separated and tested for their abilities to inhibit two alkaline serine proteinases that we previously isolated from F. culmorum. The proteins were separated by size exclusion, ion exchange, and reversed-phase-HPLC chromatographies. The purified inhibitors were identified by their molecular masses and N-terminal amino acid sequences. The proteins that inhibited the subtilisin-like Fusarium proteinase were the chymotrypsin/subtilisin (CI) inhibitors 1A, 1B, and 2A and the barley alpha-amylase/subtilisin inhibitor (BASI). Only one of the purified proteins inhibited the trypsin-like proteinase, the barley Bowman-Birk inhibitor (BBBI). No novel inhibitors were detected.

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“…DmAMP1 ( Dahlia merckii antimicrobial peptide 1) ( 31 ) and HbD2 ( 32 ) were expressed in Pichia pastoris and purified using ion-exchange chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC) as described previously ( 26 ). An inactive variant of bovine pancreatic trypsin inhibitor (BPTI) ( 25 ) (iBPTI with K15A and R17A substitutions), wild-type barley chymotrypsin inhibitor CI-1B ( 33 ), an inactive variant of CI-1B (iCI-1B with L63A substitution), At2g38870, a PR6-like protein with serine protease inhibitory activity from Arabidopsis thaliana (GenBank accession no. BT005182 ), NaPin1a, a serine protease inhibitor from N. alata (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

Synergistic Activity between Two Antifungal Proteins, the Plant Defensin NaD1 and the Bovine Pancreatic Trypsin Inhibitor

Bleackley

Dawson

McKenna

et al. 2017

mSphere

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This work describes the increased activity of a natural antifungal peptide in the presence of another antifungal peptide from a different family. This is termed antifungal synergy. Synergy is important for decreasing the amount of antifungal molecule needed to control the disease. Traditionally, naturally occurring antifungal molecules are assayed in isolation. Identification of synergistic interactions between antifungal peptides means that their activities in a complex biological system are likely to be different from what we observe when examining them individually. This study identified synergy between an antifungal peptide and a group of peptides that do not affect fungal growth in vitro. This provides the foundation for generation of transgenic plants with increased resistance to fungal disease and identification of antifungal accessory factors that enhance the activity of innate immune molecules but do not have an antifungal effect on their own.

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Expression and kinetic characterization of barley chymotrypsin inhibitors 1a and 1b

Cited by 10 publications

References 36 publications

Interaction of subtilisins with serpins

Interaction of subtilisins with serpins

Purification and Identification of Barley (Hordeum vulgare L.) Proteins That Inhibit the Alkaline Serine Proteinases of Fusarium culmorum

Synergistic Activity between Two Antifungal Proteins, the Plant Defensin NaD1 and the Bovine Pancreatic Trypsin Inhibitor

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