Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi

J. M. Ramalho-Ortigão; S. Kamhawi; M. B. Joshi; D. Reynoso; P. G. Lawyer; D. M. Dwyer; D. L. Sacks; J. G. Valenzuela

doi:10.1111/j.1365-2583.2005.00601.x

Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi

J. M. Ramalho-Ortigão, S. Kamhawi, M. B. Joshi, D. Reynoso, P. G. Lawyer, D. M. Dwyer, D. L. Sacks, J. G. Valenzuela

Research output: Contribution to journal › Article › peer-review

35 Scopus citations

Abstract

We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293-F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%.

Original language	English (US)
Pages (from-to)	703-712
Number of pages	10
Journal	Insect Molecular Biology
Volume	14
Issue number	6
DOIs	https://doi.org/10.1111/j.1365-2583.2005.00601.x
State	Published - Dec 2005
Externally published	Yes

Keywords

Chitinolytic activity
Leishmania
Peritrophic matrix
Sand fly

ASJC Scopus subject areas

Molecular Biology
Genetics
Insect Science

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10.1111/j.1365-2583.2005.00601.x

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Ramalho-Ortigão, J. M., Kamhawi, S., Joshi, M. B., Reynoso, D., Lawyer, P. G., Dwyer, D. M., Sacks, D. L., & Valenzuela, J. G. (2005). Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi. Insect Molecular Biology, 14(6), 703-712. https://doi.org/10.1111/j.1365-2583.2005.00601.x

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abstract = "We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293-F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%.",

keywords = "Chitinolytic activity, Leishmania, Peritrophic matrix, Sand fly",

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year = "2005",

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doi = "10.1111/j.1365-2583.2005.00601.x",

language = "English (US)",

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AU - Kamhawi, S.

AU - Joshi, M. B.

AU - Reynoso, D.

AU - Lawyer, P. G.

AU - Dwyer, D. M.

AU - Sacks, D. L.

AU - Valenzuela, J. G.

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N2 - We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293-F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%.

AB - We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293-F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%.

KW - Chitinolytic activity

KW - Leishmania

KW - Peritrophic matrix

KW - Sand fly

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Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi

Abstract

Keywords

ASJC Scopus subject areas

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