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Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut.
Insect Biochem Mol Biol. 2009 Nov; 39(11):782-91.IB

Abstract

Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midgut cells. The data support the view that on adapting to deal with a bacteria-rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3.

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Authors+Show Affiliations

Padilha MH
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
Pimentel AC
No affiliation info available
Ribeiro AF
No affiliation info available
Terra WR
No affiliation info available

MeSH

Amino Acid SequenceAnimalsCathepsin DCattleDigestive SystemHousefliesInsect ProteinsInsectaLysosomesMolecular Sequence DataPhylogenySequence AlignmentVertebrates

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19815068

Citation

Padilha, Marcelo H P., et al. "Sequence and Function of Lysosomal and Digestive Cathepsin D-like Proteinases of Musca Domestica Midgut." Insect Biochemistry and Molecular Biology, vol. 39, no. 11, 2009, pp. 782-91.
Padilha MH, Pimentel AC, Ribeiro AF, et al. Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut. Insect Biochem Mol Biol. 2009;39(11):782-91.
Padilha, M. H., Pimentel, A. C., Ribeiro, A. F., & Terra, W. R. (2009). Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut. Insect Biochemistry and Molecular Biology, 39(11), 782-91. https://doi.org/10.1016/j.ibmb.2009.09.003
Padilha MH, et al. Sequence and Function of Lysosomal and Digestive Cathepsin D-like Proteinases of Musca Domestica Midgut. Insect Biochem Mol Biol. 2009;39(11):782-91. PubMed PMID: 19815068.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut. AU - Padilha,Marcelo H P, AU - Pimentel,André C, AU - Ribeiro,Alberto F, AU - Terra,Walter R, Y1 - 2009/10/06/ PY - 2009/06/18/received PY - 2009/09/16/revised PY - 2009/09/17/accepted PY - 2009/10/10/entrez PY - 2009/10/10/pubmed PY - 2010/1/30/medline SP - 782 EP - 91 JF - Insect biochemistry and molecular biology JO - Insect Biochem Mol Biol VL - 39 IS - 11 N2 - Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midgut cells. The data support the view that on adapting to deal with a bacteria-rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3. SN - 1879-0240 UR - https://www.unboundmedicine.com/medline/citation/19815068/Sequence_and_function_of_lysosomal_and_digestive_cathepsin_D_like_proteinases_of_Musca_domestica_midgut_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0965-1748(09)00133-7 DB - PRIME DP - Unbound Medicine ER -