Primary structure of cathepsin D inhibitor from potatoes and its structure relationship to soybean trypsin inhibitor family

Mareš, Michael; Meloun, B.; Pavlı́k, Milan; Kostka, V.; Baudyš, Miroslav

doi:10.1016/0014-5793(89)81435-8

Cited by 107 publications

(72 citation statements)

References 27 publications

(15 reference statements)

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“…Homology of about 20% was found with trypsin inhibitors of the soybean trypsin inhibitor family 1261, aspartic proteinase inhibitors from potato [27,10] and non-inhibitory proteins miraculin (MRC) [28] and plant albumin [29]. Up to now, the tertiary structures of three proteins of this superfamily have been determined -ST1 [30], Erythrina caffra trypsin inhibitor (ETI) 1311 and wheat ~-amylase/proteinase K (subtilisin) inhibitor (WASI) 1321.…”

Section: Resultsmentioning

confidence: 99%

The primary structure of inhibitor of cysteine proteinases from potato

et al. 1993

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The complete amino acid sequence of the cysteine proteinase inhibitor from potato tubers was determined. The inhibitor is a single-chain protein having 180 amino acid residues. Its primary structure was elucidated by automatic degradation of the intact protein and sequence analysis of peptides generated by CNBr, trypsin and glycyl endopeptidase. A search through the protein sequence database showed homology to other plant proteinase inhibitors of different specificities and non-inhibitory proteins of M, around 20,000. On the basis of sequence homology, prediction of secondary structure and fold compatibility, based on a 3DlD score to the threedimensional profile of Erythrina caffra trypsin inhibitor, we suggest that the potato cysteine proteinase inhibitor belongs to the superfamily of proteins that have the same pattern of three-dimensional structure as soybean trypsin inhibitor. This superfamily would therefore include proteins that inhibit three different classes of proteinases -serine, cysteine and aspartic proteinases.Cysteine proteinase inhibitor; Amino acid sequence; Solunum tuberown; Soybean trypsin inhibitor superfamily

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Section: Resultsmentioning

confidence: 99%

The primary structure of inhibitor of cysteine proteinases from potato

et al. 1993

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“…The API preparation that we used in this study consists of several isoforms of potato cathepsin D inhibitor (PDI) that is structurally related to the soybean trypsin inhibitor family (Mares et al, 1989). It inhibits cathepsin D as well as the serine proteinases trypsin and chymotrypsin, but does not affect pepsin activity (Keilova and Tomasek, 1976a).…”

Section: Discussionmentioning

confidence: 99%

Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors

Wilhite

Elden

Brzin

et al. 2000

Insect Biochemistry and Molecular Biology

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"Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors" (2000). AbstractProteolytic activities in alfalfa weevil (Hypera postica) larval midguts have been characterized. Effects of pH, thiol activators, low-molecular weight inhibitors, and proteinase inhibitors (PIs) on general substrate hydrolysis by midgut extracts were determined. Hemoglobinolytic activity was highest in the acidic to mildly acidic pH range, but was maximal at pH 3.5. Addition of thiolactivators dithiothreitol (DTT), 2-mercaptoethanol (2-ME), or l-cysteine had little effect on hemoglobin hydrolysis at pH 3.5, but enhanced azocaseinolytic activity two to three-fold at pH 5.0. The broad cysteine PI E-64 reduced azocaseinolytic activity by 64% or 42% at pH 5 in the presence or absence of 5 mM l-cysteine, respectively. Inhibition by diazomethyl ketones, Z-Phe-Phe-CHN 2 and Z-Phe-Ala-CHN 2 , suggest that cathepsins L and B are present and comprise approximately 70% and 30% of the cysteine proteolytic activity, respectively. An aspartyl proteinase component was identified using pepstatin A, which inhibited 32% (pH 3.5, hemoglobin) and 50% (pH 5, azocasein) of total proteolytic activity. This activity was completely inhibited by an aspartyl proteinase inhibitor from potato (API), and is consistent with the action of a cathepsin D-like enzyme. Hence, genes encoding PIs with specificity toward cathepsins L, B and D could potentially be effective for control of alfalfa weevil using transgenic plants.

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“…However, it is only recently that Kunitz-type inhibitors from potato tubers have been described in detail. For example, a novel inhibitor of cathepsin D, an acid proteinase, has been isolated and characterized from potato tubers (10), and the amino acid sequence of this protein (and another closely related inhibitor, 16) is homologous to the Kunitz family of proteinase inhibitors. Also, the sequence of an abundant 22 kD protein from potato tubers has homology to Kunitz trypsin inhibitors (19,20).…”

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confidence: 99%

Two Kunitz-Type Proteinase Inhibitors from Potato Tubers

Walsh

Twitchell²

1991

Plant Physiol.

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Two proteinase inhibitors have been isolated from tubers of potato (Solanum tuberosum). Based on N-terminal amino acid sequence homologies, they are members of the Kunitz family of proteinase inhibitors. Potato Kunitz inhibitor-1 (molecular weight 19,500, isoelectric point 6.9) is a potent inhibitor of the animal pancreatic proteinase trypsin, and its amino terminus has significant homology to a recently characterized cathepsin D Kunitz inhibitor from potato tubers (Mares et a!. [1989] (19,20). In this paper, we describe two serine proteinase inhibitors from potato tubers that have differing proteinase inhibitory profiles but are both Kunitztype inhibitors based on amino-terminal sequence homologies. One closely resembles the previously described cathepsin D inhibitor (10). The other is an inhibitor of the microbial proteinase subtilisin and appears to be the same as the abundant 22 kD protein isolated by Suh et al. ( 19). This is the first Kunitz-type inhibitor of a microbial proteinase to be identified in a source other than from small grains. MATERIALS AND METHODSPotato (Solanum tuberosuim) tubers (var Russet Burbank) were purchased from a local market. Bovine trypsin, bovine Mono S 5/5 column equilibrated in 25 mm sodium acetate (pH 5.0) and using a 0 to 0.5 M NaCl gradient over 30 min with a flow rate of 1 mL/min. Reverse phase HPLC was performed with a Vydac C4 4.5 x 300 mm column equilibrated in 0. 1% TFA and eluted with a gradient of 0 to 35% acetonitrile over 8 min, then 35 to 65% acetonitrile over 30 min at a flow rate of 1 mL/min. Peaks were collected and lyophilized prior to SDS-PAGE analysis and amino terminal sequencing.15

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Primary structure of cathepsin D inhibitor from potatoes and its structure relationship to soybean trypsin inhibitor family

Cited by 107 publications

References 27 publications

The primary structure of inhibitor of cysteine proteinases from potato

The primary structure of inhibitor of cysteine proteinases from potato

Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors

Two Kunitz-Type Proteinase Inhibitors from Potato Tubers

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