Distribution of Carboxypeptidase Isoinhibitors in the Potato Plant

Hass, George M.; Derr, Judith E.; Makus, D. J.

doi:10.1104/pp.64.6.1029

Cited by 7 publications

(3 citation statements)

References 16 publications

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“…Isoinhibitors IIb and III could also derive from different processing of this same precursor protein [30]. Taken together, forms II and III represent 95% of the total inhibitor found in potato leaf [36]. The cDNA for the isoinhibitor I, the minor form in leaf, which differs in two amino acids from form II, was not found in our screening of the cDNA library.…”

Section: Discussionmentioning

confidence: 82%

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹

et al. 1998

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A partial cDNA clone for the potato wound-inducible metallocarboxypeptidase inhibitor (PCI) was isolated from a cDNA library constructed from mRNA of abscisic acid (ABA)-treated potato leaves. The full 5P region of the cDNA was obtained through a RACE-PCR protocol. PCI mRNA encodes a precursor polypeptide which comprises a 29 residue N-terminal signal peptide, a 27 residue N-terminal pro-region, the 39 residue mature PCI protein, and a 7 residue C-terminal extension. Northern blot analysis demonstrates that the PCI gene is transcriptionally activated by wounding, and wound signaling can be induced by ABA and jasmonic acid. Subcellular localization of the protein was investigated by immunocytochemistry and electron microscopy, showing that PCI accumulates within the vacuole. A partial PCI precursor form, comprising the mature protein and the C-terminal extension, has been expressed in Escherichia coli and characterized. Its inability to inhibit carboxypeptidases, and stability to carboxypeptidase digestion, suggest that the C-terminal pro-domain may have, besides a probable vacuolar sorting function, a role in modulation of the inhibitory activity of PCI.z 1998 Federation of European Biochemical Societies.

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Section: Discussionmentioning

confidence: 82%

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹

et al. 1998

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“…Potato carboxypeptidase inhibitor (PCI) competitively inhibits several metallocarboxypeptidases with a K i in the nanomolar range (Hass and Derr, 1979). A similar inhibitory activity towards metallocarboxypeptidase A (CPA) has been shown with tomato TCMP‐1 extracted from wound‐induced leaves (Diez‐Diaz et al ., 2004).…”

Section: Resultsmentioning

confidence: 99%

Anti‐angiogenic effects of two cystine‐knot miniproteins from tomato fruit

Cavallini

Trettene

Degan

et al. 2011

British J Pharmacology

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BACKGROUND AND PURPOSECystine-knot miniproteins are characterized by a similar molecular structure. Some cystine-knot miniproteins display therapeutically useful biological activities, as antithrombotic agents or tumour growth inhibitors. A critical event in the progression of tumours is the formation of new blood vessels. The aim of this work was to test two tomato cystine-knot miniproteins for their effects on endothelial cell proliferation and angiogenesis in vitro. EXPERIMENTAL APPROACHTwo tomato cystine-knot miniproteins (TCMPs) were expressed and purified either as recombinant or as native proteins from tomato fruits. The Matrigel assay was used to investigate the effects of TCMPs on in vitro angiogenesis. Viability and proliferation of endothelial cells were tested. Extracellular signal-regulated kinase (ERK)1/2 phosphorylation was assayed in either HUVEC or A431 epidermal growth factor receptor (EGFR)-overexpressing cells treated with TCMPs. EGFR phosphorylation was tested in A431 cells. KEY RESULTSBoth recombinant and native TCMPs inhibited in vitro angiogenesis of HUVEC cells at concentrations of 15-100 nM. The anti-angiogenic effect of TCMPs was associated with the inhibition of ERK phosphorylation. The two miniproteins did not alter the viability and proliferation of the endothelial cells. CONCLUSIONS AND IMPLICATIONSThe anti-angiogenetic properties of TCMPs are of potential pharmacological interest because they are common and natural components of the human diet, they possess low toxicity, they are active at submicromolar concentrations, they share a common molecular structure that can be used as a molecular platform for the design of molecules with enhanced biological activity. AbbreviationsADAM, a disintegrin and metalloprotease; CPA, bovine carboxypeptidase A; EGF, epidermal growth factor; EGFR, epidermal growth factor receptor; ERK, extracellular signal-regulated kinase; HUVEC, human umbilical vein endothelial cell; MAPK, mitogen-activated protein kinase; MMP, matrix metalloproteinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PCI, potato metallocarboxypeptidase inhibitor; TCMP, tomato cystine-knot miniprotein

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“…Hippuryl-l-arginine was used as a specific substrate to determine the level of carboxypeptidase B activity (EC 3.4.17.2) ( Hass and Derr 1979 ). The reaction mixture consisted of 2.9 ml of 1 mM hippuryl-l-arginine in 25 mM HCl buffer and 0.1 ml of gut homogenate.…”

Section: Methodsmentioning

confidence: 99%

Midgut Protease Activity During Larval Development of Anastrepha obliqua (Diptera: Tephritidae) Fed With Natural and Artificial Diet

Rivera-Ciprian

Aceituno‐Medina

Guillén-Navarro

et al. 2017

Journal of Insect Science

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In this study, we examined the activity of two serine proteases (chymotrypsin and trypsin) and two metalloproteases (carboxypeptidases A and B) during larval development in Anastrepha obliqua fed natural (mango fruit) and artificial (formulation used in mass-rearing) diets. Proteolytic activity of chymotrypsin, trypsin, carboxypeptidase A, and carboxypeptidase B was detected in the midgut of different instars of A. obliqua and was strongly affected by the pH and diet type. The protein content of the natural and artificial diets was similar. Enzymatic activity was higher in the midgut of the larvae fed the natural diet than in larvae fed the artificial diet. The activity of the endopeptidases (chymotrypsin and trypsin) was lower than those of the exopeptidases (carboxypeptidases A and B). The pH of the midgut varied from acidic to neutral. The results indicate that in the midgut of the larvae reared on both types of diet, the level of carboxypeptidase activity was approximately 100-fold greater than the level of chymotrypsin activity and 10,000-fold greater than the level of trypsin. In conclusion, carboxypeptidase A and B are the main proteases involved in the digestion of proteins in the larvae of A. obliqua. The natural diet showed a high bioaccessibility. A clear tendency to express high activities of chymotrypsin and trypsin was observed by the third instar. Our research contributes to the planning and development of novel bioaccessibility assays to understand the nutrition processing of A. obliqua larvae under mass-rearing conditions for sterile insect technique.

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Distribution of Carboxypeptidase Isoinhibitors in the Potato Plant

Cited by 7 publications

References 16 publications

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹

Anti‐angiogenic effects of two cystine‐knot miniproteins from tomato fruit

Midgut Protease Activity During Larval Development of Anastrepha obliqua (Diptera: Tephritidae) Fed With Natural and Artificial Diet

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Distribution of Carboxypeptidase Isoinhibitors in the Potato Plant

Cited by 7 publications

References 16 publications

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato1

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato1

Anti‐angiogenic effects of two cystine‐knot miniproteins from tomato fruit

Midgut Protease Activity During Larval Development of Anastrepha obliqua (Diptera: Tephritidae) Fed With Natural and Artificial Diet

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Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹

Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato¹