title: Induction of leucine aminopeptidase (LAP) like activity with wounding and methyl jasmonate in pigeonpea (Cajanas cajan) suggests the role of these enzymes in plant defense in leguminosae.
abstract: Aminopeptidases are ubiquitous in nature and their activities have been identified in several plant species. Leucine aminopeptidases (LAPs) are predominantly studied in solanaceous plants and are induced in response to wounding, herbivory and methyl jasmonate (MeJA). The functions of plant aminopeptidases are still under discussion and it is likely that the different classes play various roles. In the present study we report the local and systemic induction of LAP-like activity upon mechanical wounding and MeJA treatment. Two proteins with LAP-like activity were detected in pigeonpea leaves. They were designated as AP1 and AP2. AP1 activity was significantly induced upon wounding and application of MeJA. The estimated molecular masses of AP1 and AP2 were ∼ 60 and 41 kDa respectively in SDS-PAGE. The pH optimum for LAP-like activity in control leaf extracts was found to be neutral (pH 7.0) however the enzymes showed highest activity at alkaline pH (pH 9.0) in the leaf extracts of treated plants. The temperature optimum for LAP-like activity was around 40-50 °C. The enzymes were strongly inhibited by 1, 10 phenanthroline and bestatin. Heavy metal ions and EDTA inhibited LAP-like activities, whereas Mn(+2) and Mg(+2) activated the enzyme activities. Beside LpNA (33.5 U/mg/min) pigeonpea LAP-like enzymes also cleaved ApNA (15 U/mg/min) but were unable to cleave VpNA. Total proteolytic activity was also observed to be induced in treated plants. LAP-like activity was increased upto 19.5 fold after gel filtration chromatography. Results suggest that these enzymes may have functional defensive role in pigeonpea.
authors.affiliation: Department of Biochemistry, Dr Babasaheb Ambedkar Marathwada University, Aurangabad 431004, Maharashtra, India.
authors.forename: Purushottam R
authors.initials: PR
authors.lastname: Lomate
authors.forename: Vandana K
authors.initials: VK
authors.lastname: Hivrale
mesh_terms: D000085:Acetates / Q000494:pharmacology*; D000222:Adaptation, Physiological; D036561:Cajanus / Q000187:drug effects / Q000201:enzymology*; D003517:Cyclopentanes / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007931:Leucyl Aminopeptidase / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D054883:Oxylipins / Q000494:pharmacology*; D010935:Plant Diseases; D010937:Plant Growth Regulators / Q000494:pharmacology*; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology; D010940:Plant Proteins / Q000378:metabolism*; D013696:Temperature
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D004791:Enzyme Inhibitors; D054883:Oxylipins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D008345:Manganese; C072239:methyl jasmonate; D007931:Leucyl Aminopeptidase; D008274:Magnesium