The amino acid sequence of a novel inhibitor of cathepsin D from potato

Ritonja, Anka; Križaj, Igor; Meško, Pika; Kopitar, M.; Lučovnik, P.; Štrukelj, Borut; Pungerčar, Jože; Buttle, David J.; Barrett, Alan J.; Türk, Vito

doi:10.1016/0014-5793(90)80275-n

Cited by 64 publications

(48 citation statements)

References 13 publications

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“…Oryzacystatin I (OCI) (Abe et al, 1987) was expressed as s glutathione S-transferase (GST) fusion protein in Escherichia coli strain BL21 (Stratagene) as previously described (Michaud et al, 1994). Aspartyl proteinase inhibitor (API) was purified from potato as previously described (Kreft et al, 1997) and consisted of several isoforms (Ritonja et al, 1990;Barlic-Maganja et al, 1992;Strukelj et al 1992Strukelj et al , 1995Kreft et al, 1997) of potato cathepsin D inhibitor (PDI) (Keilova and Tomasek, 1976b).…”

Section: Methodsmentioning

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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Section: Methodsmentioning

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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“…The amino-terminal sequence of PKI-1 has strong homology to the sequences of two closely related cathepsin-D inhibitors (10,16), with identity between residues 21 to 42. These cathepsin D inhibitors, like PKI-1, possess significant inhibitory activity against trypsin.…”

Section: Assays Of Proteinase Inhibitionmentioning

confidence: 99%

“…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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“…Within the last few years, a large number of protein inhibitors of cysteine proteinases have also been discovered and characterized [3,4]. In contrast, only a few protein inhibitors directed towards metallo-proteinases (TIMP and PCI, see [5,61) or aspartyl proteinases (see [7][8][9]) are known to date. The azmacroglobulin family presents an exception, as these proteins can inhibit all of these proteinases according to a 'molecular trap' mechanism by virtue of a promiscuous 'bait region' (see [IOI).…”

mentioning

confidence: 99%

Natural protein proteinase inhibitors and their interaction with proteinases

Bode

Huber

1992

European Journal of Biochemistry

1,010

624

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The substrate-like 'canonical' inhibition by the 'small' serine proteinase inhibitors and the productlike inhibition by the carboxypeptidase inhibitor have provided the only atomic models of protein inhibitor -proteinase interactions for about 15 years. The recently published structures of cystatin/ stefin -papain complexes and of hirudin -thrombin complexes reveal novel non-substrate-like interactions, In addition, the structure of pro-carboxypeptidase showes a mode of inactivation which bears resemblance to proteinase/protein inhibitor systems. Considerable progress in understanding the transition between native and cleaved states of the serpins has also been made by several recent structural studies.Proteinase inhibitors are important tools of nature for regulating the proteolytic activity of their target proteinases, for blocking these in emergency cases, or for signaling receptor interactions or clearance. Endogenous inhibitors appear to be always proteins; small non-proteinaceous inhibitors which impair the proteolytic activity of host proteinases are produced in microorganisms.The number of proteinaceous proteinase inhibitors isolated and identified so far is extremely large. In a now classical review paper, Laskowski and Kato [2] introduced for the first time a rational nomenclature by grouping these diverse inhibitors into distinct protein families. In the meantime, this list of families has considerably expanded with the advent of many new inhibitor species, and is still growing.The majority of protein inhibitors known and characterized so far are directed towards serine proteinases. Within the last few years, a large number of protein inhibitors of cysteine proteinases have also been discovered and characterized [3,4]. In contrast, only a few protein inhibitors directed towards metallo-proteinases (TIMP and PCI, see [5,61) or aspartyl proteinases (see [7][8][9]) are known to date. The azmacroglobulin family presents an exception, as these proteins can inhibit all of these proteinases according to a 'molecular trap' mechanism by virtue of a promiscuous 'bait region' (see [IOI).Until recently, X-ray crystal structures of only a few serine proteinase inhibitors, one carboxypeptidase inhibitor, and some of their complexes with cognate proteinases were available. The X-ray crystal structures of protein inhibitors published up to 1985 have been reviewed by Read and James A new aspect is provided by inhibitor structures elucidated by two-dimensional NMR methods (see [22, 231 for reviews). These data are often complementary to X-ray data, but are restricted to isolated inhibitors of relatively small molecular mass; until now, no NMR structure of a protein inhibitor has been reported for whch there is no X-ray structure available.In this review, we shall attempt to illuminate the characteristic structural properties conferring inhibitory activity to proteins. Nature has used diverse approaches to achieve proteinase inhibition. This is particularly well illustrated by some more recently published structures. In...

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The amino acid sequence of a novel inhibitor of cathepsin D from potato

Cited by 64 publications

References 13 publications

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

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

Natural protein proteinase inhibitors and their interaction with proteinases

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