Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae
Само за регистроване кориснике
2008
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 th...at 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.
Кључне речи:
Cerambycid beetle / Isoform / Leucyl aminopeptidase / Midgut / Morimus funereus / Xylophagous larvaeИзвор:
Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 2008, 149, 3, 454-462Издавач:
- Elsevier Science Inc, New York
Финансирање / пројекти:
- Интеракције природних производа и њихових аналога са протеинима и нуклеинским киселинама (RS-MESTD-MPN2006-2010-142026)
DOI: 10.1016/j.cbpb.2007.11.006
ISSN: 1096-4959
PubMed: 18155948
WoS: 000253576300007
Scopus: 2-s2.0-39049120322
Институција/група
IHTMTY - JOUR AU - Božić, Nataša AU - Ivanović, J. AU - Nenadović, V. AU - Bergström, J. AU - Larsson, T. AU - Vujčić, Zoran PY - 2008 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/463 AB - 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. PB - Elsevier Science Inc, New York T2 - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology T1 - Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae VL - 149 IS - 3 SP - 454 EP - 462 DO - 10.1016/j.cbpb.2007.11.006 ER -
@article{ author = "Božić, Nataša and Ivanović, J. and Nenadović, V. and Bergström, J. and Larsson, T. and Vujčić, Zoran", year = "2008", 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.", publisher = "Elsevier Science Inc, New York", journal = "Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology", title = "Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae", volume = "149", number = "3", pages = "454-462", doi = "10.1016/j.cbpb.2007.11.006" }
Božić, N., Ivanović, J., Nenadović, V., Bergström, J., Larsson, T.,& Vujčić, Z.. (2008). Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae. in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology Elsevier Science Inc, New York., 149(3), 454-462. https://doi.org/10.1016/j.cbpb.2007.11.006
Božić N, Ivanović J, Nenadović V, Bergström J, Larsson T, Vujčić Z. Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae. in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology. 2008;149(3):454-462. doi:10.1016/j.cbpb.2007.11.006 .
Božić, Nataša, Ivanović, J., Nenadović, V., Bergström, J., Larsson, T., Vujčić, Zoran, "Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae" in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 149, no. 3 (2008):454-462, https://doi.org/10.1016/j.cbpb.2007.11.006 . .