Characterisation of aminopeptidase activity in scab mites, Psoroptes spp.

A. J. Nisbet, Peter Francis Billingsley

    Research output: Contribution to journalArticle

    13 Citations (Scopus)

    Abstract

    Soluble and membrane-bound aminopeptidase activities were demonstrated in extracts of P. cuniculi (Delafond). Leucine amino-peptidase (LAP) activity in the soluble fraction of P. cuniculi extracts displayed substrate preference for amino acid derivatives with terminal leucine and methionine over those with acidic, basic or heterocyclic groups. P. cuniculi LAP was inhibited by leucinethiol (IC50 = 1.4 +/- 0.4 nM), bestatin (IC50 = 3.9 +/- 1.7 muM), Arphamenine A (IC50 = 0.37 +/- 0.03 mM) the chelating agent 1,10-phenanthroline (IC50 = 2.3 +/- 0.5 mM), Zn2+, Cu2+ Ni2+, and Co2+, and activated by Mn2+ and Mg2+. The LAP activity was visualised as a single major band after electrophoresis on native gels and eluted from a size exclusion column as a single major peak representing a molecular mass range of 85-116 kDa.

    Degenerate oligonucleotide primers were used to amplify short fragments of genomic DNA containing nucleotide sequence coding for the cation-binding motifs of the co-catalytic Zn2+ binding domains of dizinc leucine aminopeptidases in both P. cuniculi and P.ovis (Hering). The major soluble aminopeptidase from these mites therefore displays most of the characteristics associated with typical cytosolic leucine aminopeptidases belonging to the M17 family of metalloproteinases. (C) 2002 Elsevier Science Ltd. All rights reserved.

    Original languageEnglish
    Pages (from-to)1123-1131
    Number of pages8
    JournalInsect Biochemistry and Molecular Biology
    Volume32
    DOIs
    Publication statusPublished - 2002

    Keywords

    • Psoroptes
    • aminopeptidase
    • LAP
    • digestion
    • psoroptic mange
    • BRUSH-BORDER MEMBRANE
    • LEUCINE AMINOPEPTIDASE
    • HYDROLYTIC ENZYMES
    • BLOOD DIGESTION
    • BOVINE LENS
    • MIDGUT
    • SHEEP
    • MECHANISM
    • PROTEINASES
    • INHIBITORS

    Cite this

    Characterisation of aminopeptidase activity in scab mites, Psoroptes spp.. / Nisbet, A. J.; Billingsley, Peter Francis.

    In: Insect Biochemistry and Molecular Biology, Vol. 32, 2002, p. 1123-1131.

    Research output: Contribution to journalArticle

    Nisbet, A. J. ; Billingsley, Peter Francis. / Characterisation of aminopeptidase activity in scab mites, Psoroptes spp. In: Insect Biochemistry and Molecular Biology. 2002 ; Vol. 32. pp. 1123-1131.
    @article{dff450c32e2f42f0abd3b2acffcdb9de,
    title = "Characterisation of aminopeptidase activity in scab mites, Psoroptes spp.",
    abstract = "Soluble and membrane-bound aminopeptidase activities were demonstrated in extracts of P. cuniculi (Delafond). Leucine amino-peptidase (LAP) activity in the soluble fraction of P. cuniculi extracts displayed substrate preference for amino acid derivatives with terminal leucine and methionine over those with acidic, basic or heterocyclic groups. P. cuniculi LAP was inhibited by leucinethiol (IC50 = 1.4 +/- 0.4 nM), bestatin (IC50 = 3.9 +/- 1.7 muM), Arphamenine A (IC50 = 0.37 +/- 0.03 mM) the chelating agent 1,10-phenanthroline (IC50 = 2.3 +/- 0.5 mM), Zn2+, Cu2+ Ni2+, and Co2+, and activated by Mn2+ and Mg2+. The LAP activity was visualised as a single major band after electrophoresis on native gels and eluted from a size exclusion column as a single major peak representing a molecular mass range of 85-116 kDa.Degenerate oligonucleotide primers were used to amplify short fragments of genomic DNA containing nucleotide sequence coding for the cation-binding motifs of the co-catalytic Zn2+ binding domains of dizinc leucine aminopeptidases in both P. cuniculi and P.ovis (Hering). The major soluble aminopeptidase from these mites therefore displays most of the characteristics associated with typical cytosolic leucine aminopeptidases belonging to the M17 family of metalloproteinases. (C) 2002 Elsevier Science Ltd. All rights reserved.",
    keywords = "Psoroptes, aminopeptidase, LAP, digestion, psoroptic mange, BRUSH-BORDER MEMBRANE, LEUCINE AMINOPEPTIDASE, HYDROLYTIC ENZYMES, BLOOD DIGESTION, BOVINE LENS, MIDGUT, SHEEP, MECHANISM, PROTEINASES, INHIBITORS",
    author = "Nisbet, {A. J.} and Billingsley, {Peter Francis}",
    year = "2002",
    doi = "10.1016/S0965-1748(02)00048-6",
    language = "English",
    volume = "32",
    pages = "1123--1131",
    journal = "Insect Biochemistry and Molecular Biology",
    issn = "0965-1748",
    publisher = "Elsevier Limited",

    }

    TY - JOUR

    T1 - Characterisation of aminopeptidase activity in scab mites, Psoroptes spp.

    AU - Nisbet, A. J.

    AU - Billingsley, Peter Francis

    PY - 2002

    Y1 - 2002

    N2 - Soluble and membrane-bound aminopeptidase activities were demonstrated in extracts of P. cuniculi (Delafond). Leucine amino-peptidase (LAP) activity in the soluble fraction of P. cuniculi extracts displayed substrate preference for amino acid derivatives with terminal leucine and methionine over those with acidic, basic or heterocyclic groups. P. cuniculi LAP was inhibited by leucinethiol (IC50 = 1.4 +/- 0.4 nM), bestatin (IC50 = 3.9 +/- 1.7 muM), Arphamenine A (IC50 = 0.37 +/- 0.03 mM) the chelating agent 1,10-phenanthroline (IC50 = 2.3 +/- 0.5 mM), Zn2+, Cu2+ Ni2+, and Co2+, and activated by Mn2+ and Mg2+. The LAP activity was visualised as a single major band after electrophoresis on native gels and eluted from a size exclusion column as a single major peak representing a molecular mass range of 85-116 kDa.Degenerate oligonucleotide primers were used to amplify short fragments of genomic DNA containing nucleotide sequence coding for the cation-binding motifs of the co-catalytic Zn2+ binding domains of dizinc leucine aminopeptidases in both P. cuniculi and P.ovis (Hering). The major soluble aminopeptidase from these mites therefore displays most of the characteristics associated with typical cytosolic leucine aminopeptidases belonging to the M17 family of metalloproteinases. (C) 2002 Elsevier Science Ltd. All rights reserved.

    AB - Soluble and membrane-bound aminopeptidase activities were demonstrated in extracts of P. cuniculi (Delafond). Leucine amino-peptidase (LAP) activity in the soluble fraction of P. cuniculi extracts displayed substrate preference for amino acid derivatives with terminal leucine and methionine over those with acidic, basic or heterocyclic groups. P. cuniculi LAP was inhibited by leucinethiol (IC50 = 1.4 +/- 0.4 nM), bestatin (IC50 = 3.9 +/- 1.7 muM), Arphamenine A (IC50 = 0.37 +/- 0.03 mM) the chelating agent 1,10-phenanthroline (IC50 = 2.3 +/- 0.5 mM), Zn2+, Cu2+ Ni2+, and Co2+, and activated by Mn2+ and Mg2+. The LAP activity was visualised as a single major band after electrophoresis on native gels and eluted from a size exclusion column as a single major peak representing a molecular mass range of 85-116 kDa.Degenerate oligonucleotide primers were used to amplify short fragments of genomic DNA containing nucleotide sequence coding for the cation-binding motifs of the co-catalytic Zn2+ binding domains of dizinc leucine aminopeptidases in both P. cuniculi and P.ovis (Hering). The major soluble aminopeptidase from these mites therefore displays most of the characteristics associated with typical cytosolic leucine aminopeptidases belonging to the M17 family of metalloproteinases. (C) 2002 Elsevier Science Ltd. All rights reserved.

    KW - Psoroptes

    KW - aminopeptidase

    KW - LAP

    KW - digestion

    KW - psoroptic mange

    KW - BRUSH-BORDER MEMBRANE

    KW - LEUCINE AMINOPEPTIDASE

    KW - HYDROLYTIC ENZYMES

    KW - BLOOD DIGESTION

    KW - BOVINE LENS

    KW - MIDGUT

    KW - SHEEP

    KW - MECHANISM

    KW - PROTEINASES

    KW - INHIBITORS

    U2 - 10.1016/S0965-1748(02)00048-6

    DO - 10.1016/S0965-1748(02)00048-6

    M3 - Article

    VL - 32

    SP - 1123

    EP - 1131

    JO - Insect Biochemistry and Molecular Biology

    JF - Insect Biochemistry and Molecular Biology

    SN - 0965-1748

    ER -