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Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae
dc.creator | Božić, Nataša | |
dc.creator | Ivanović, J. | |
dc.creator | Nenadović, V. | |
dc.creator | Bergström, J. | |
dc.creator | Larsson, T. | |
dc.creator | Vujčić, Zoran | |
dc.date.accessioned | 2019-01-30T17:18:49Z | |
dc.date.available | 2019-01-30T17:18:49Z | |
dc.date.issued | 2008 | |
dc.identifier.issn | 1096-4959 | |
dc.identifier.uri | https://cer.ihtm.bg.ac.rs/handle/123456789/463 | |
dc.description.abstract | The major leucyl aminopeptidase (LAP) from the midgut of Morimus funereus larvae was purified and characterised. Specific LAP activity was increased 292-fold by purification of the crude midgut extract. The purified enzyme had a pH optimum of 7.5 (optimum pH range 7.0-8.5) and preferentially hydrolysed p-nitroanilides containing hydrophobic amino acids in the active site, with the highest Vmax / KM ratio for leucine-p-nitroanilide (LpNA). Among a number of inhibitors tested, the most efficient were 1,10-phenanthroline having a Ki value of 0.12 mM and cysteine with Ki value of 0.31 mM, while EGTA stimulated LAP activity. Zn2+, Mg2+ and Mn2+ all showed bi-modal effects on LAP activity (activated at low concentrations and inhibited at high concentrations). The purified LAP (after gel filtration on Superose 6 column) had molecular mass of 400 kDa with an isoelectric point of 6.2. Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed one band of 67 kDa, suggesting that the enzyme is a hexamer. Six peptide sequences from protein band were obtained using ESI/MS-MS analysis. Comparison of the obtained peptide sequences with the EMBL-EBI sequence analysis toolbox and the BLASTP database showed a high degree of identity with other insect aminopeptidases. | en |
dc.publisher | Elsevier Science Inc, New York | |
dc.relation | info:eu-repo/grantAgreement/MESTD/MPN2006-2010/142026/RS// | |
dc.rights | restrictedAccess | |
dc.source | Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology | |
dc.subject | Cerambycid beetle | en |
dc.subject | Isoform | en |
dc.subject | Leucyl aminopeptidase | en |
dc.subject | Midgut | en |
dc.subject | Morimus funereus | en |
dc.subject | Xylophagous larvae | en |
dc.title | Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae | en |
dc.type | article | |
dc.rights.license | ARR | |
dcterms.abstract | Ивановић, Ј.; Божић, Наташа; Бергстрöм, Ј.; Вујчић, З.; Ларссон, Т.; Ненадовић, В.; | |
dc.citation.volume | 149 | |
dc.citation.issue | 3 | |
dc.citation.spage | 454 | |
dc.citation.epage | 462 | |
dc.citation.other | 149(3): 454-462 | |
dc.citation.rank | M22 | |
dc.identifier.pmid | 18155948 | |
dc.identifier.doi | 10.1016/j.cbpb.2007.11.006 | |
dc.identifier.scopus | 2-s2.0-39049120322 | |
dc.identifier.wos | 000253576300007 | |
dc.type.version | publishedVersion |