Herein we describe the cloning and characterization of a cDNA encoding an aspartic proteinase from the root-knot nematode Meloidogyne incognita. Using PCR techniques, a 1471-bp cDNA fragment encoding a cathepsin D-like (Mi-asp1) transcript was isolated from second-stage larvae mRNA. Its predicted amino acid sequence comprises a pro-region of 71 amino acid residues and a mature protease of 378 amino acid residues with a predicted molecular mass of 41.502kDa. Protein sequence comparisons of Mi-asp1 with GenBank (DQ360827) sequences showed 59-71% identity with nematode-specific cathepsin D-like aspartic proteinases. Southern blot analysis, RT-PCR amplification and EST mining suggest the existence of a developmentally expressed gene family encoding aspartic proteinases in M. incognita. Mi-asp1 may represent a potential target for molecular intervention for the purposes of plant-parasitic nematode control.
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http://dx.doi.org/10.1016/j.exppara.2008.09.017 | DOI Listing |
Pestic Biochem Physiol
January 2022
Department of Zoology, University of Madras, Guindy Campus, Chennai 600 025, India. Electronic address:
The pulse beetle Callosobruchus maculatus causes potential damage to legume crops by infesting the seeds, leading to a reduction of total protein content. Arcelin found in the wild accessions of the common bean, is an insecticidal protein that has the potency to hamper the metabolism of the bruchid beetle. The arcelin gene from the wild accession of Phaseolus lunatus was isolated and the ORF encoding 158 amino acids was cloned in pET-45b (+) vector.
View Article and Find Full Text PDFGenome
December 2021
Anhui Province Key Laboratory of Aquaculture & Stock Enhancement, Fisher Institute of Anhui Academy of Agricultural Sciences, Hefei, 230031, China.
Aspartic proteinases are one of the four families of proteinase enzymes that are widely present in living organisms. They are involved in various physiological events, such as protein degradation, development, and host defense. However, the characterization and functional roles of aspartic proteinases remain to be elucidated in crustaceans.
View Article and Find Full Text PDFmSphere
February 2020
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA
The lysosome-like vacuolar compartment (VAC) is a major site of proteolysis in the intracellular parasite Previous studies have shown that genetic ablation of a VAC-residing cysteine protease, cathepsin protease L (CPL), resulted in the accumulation of undigested protein in the VAC and loss of parasite viability during the chronic stage of infection. However, since the maturation of another VAC localizing protease, cathepsin protease B (CPB), is dependent on CPL, it remained unknown whether these defects result directly from ablation of CPL or indirectly from a lack of CPB maturation. Likewise, although a previously described cathepsin D-like aspartyl protease 1 (ASP1) could also play a role in proteolysis, its definitive residence and function in the endolysosomal system were not well defined.
View Article and Find Full Text PDFProtein Expr Purif
March 2020
Laboratório de Bioquímica Experimental e Computacional de Fármacos, LaBECFar, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Avenida Brasil 4365, CEP 21040-360, Rio de Janeiro, RJ, Brazil. Electronic address:
Schistosomes express a variety of aspartyl proteases (APs) with distinct roles in the helminth pathophysiology, among which degradation of host haemoglobin is key, since it is the main amino acid source for these parasites. A cathepsin D-like AP from Schistosoma mansoni (SmCD1) has been used as a model enzyme for vaccine and drug development studies in schistosomes and yet a reliable expression system for readily producing the recombinant enzyme in high yield has not been reported. To contribute to further advancing the knowledge about this valuable antischistosomal target, we developed a transient expression system in HEK 293T mammalian cells and performed a biochemical and biophysical characterization of the recombinant enzyme (rSmCD1).
View Article and Find Full Text PDFBiomolecules
June 2019
School of Food Science and Pharmacy, Zhejiang Ocean University, Zhoushan 316000, China.
A protease from neon flying squid () viscera (SVCE3(f)) was partially purified by isoelectric solubilization/precipitation combined with ultra-membrane filtration (ISP-UMF). Two protein bands of 45 and 27 KDa were determined by SDS-PAGE assay. The protease characteristic of the protein band of 45 KDa was confirmed using casein zymography analysis.
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