@article {466,
	title = {Characterisation of functional and insecticidal properties of a recombinant cathepsin L-like proteinase from flesh fly (Sarcophaga peregrina), which plays a role in differentiation of imaginal discs},
	volume = {37},
	year = {Submitted},
	note = {08-10-200906-02-2012NormalSchool of Biological and Biomedical Sciences, Durham University, South Road, Durham, UK.enghttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=17517336},
	pages = {589 - 600},
	abstract = {ScathL is a cathepsin L-like cysteine proteinase from Sarcophaga peregrina (flesh fly), which is involved in differentiation of imaginal discs, through proteolysis of components of basement membranes. An expression system based on the methylotrophic yeast Pichia pastoris was used to produce recombinant ScathL. Although the expression construct contained the full proprotein coding sequence for ScathL, the proprotein was only detected in culture supernatant at early stages of expression by Western blotting. The purified recombinant protein contained only a polypeptide similar to mature ScathL, as a result of autocatalytic processing. After activation by reducing agents, the enzyme hydrolysed the cathepsin L substrate Z-Phe-Arg-AMC, with optimal activity at pH 5.5. ScathL showed decreasing activity with increasing ionic strength above 0.3M NaCl, and lost activity irreversibly at pH \&gt; or = 7.5. The enzyme showed limited activity towards protein substrates, digesting only to large fragments. ScathL was insecticidal towards larvae of the tomato moth, Lacanobia oleracera, following injection into the haemolymph. It caused melanisation, although no evidence of extensive proteolysis in haemolymph or gut was observed. Production of a inactive mutant form of ScathL showed that enzyme activity was necessary for the complete proprotein processing observed during production as a recombinant protein, and for insecticidal activity.},
	keywords = {Amino Acid Sequence, Animals, Cysteine Endopeptidases/genetics/isolation \&amp, development, Diptera/*enzymology/growth \&amp, Enzyme Activation, Hemolymph/metabolism, Hydrogen-Ion Concentration, Insecticides/pharmacology, Larva/drug effects, Lepidoptera/drug effects, Mutation, Pichia/genetics, purification/*metabolism/pharmacology, purification/metabolism/pharmacology, Recombinant Proteins/genetics/isolation \&amp, Salts/metabolism},
	author = {Philip, J. M. and Fitches, E. and Harrison, R. L. and Bonning, B. and Gatehouse, J. A.}
}
@article {365,
	title = {Insecticidal activity of a basement membrane-degrading protease against Heliothis virescens (Fabricius) and Acyrthosiphon pisum (Harris)},
	volume = {54},
	year = {Submitted},
	note = {08-10-200906-02-2012NormalDepartment of Entomology, Iowa State University, 418 Science II, Ames, IA 50011-3222, USA.enghttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=18395222},
	pages = {777 - 789},
	abstract = {ScathL is a cathepsin L-like cysteine protease derived from the flesh fly Sarcophaga peregrina that functions in basement membrane (BM) remodeling during insect development. A recombinant baculovirus expressing ScathL (AcMLF9.ScathL) kills larvae of the tobacco budworm, Heliothis virescens, significantly faster than the wild-type virus. Here, we show that the occurrence of larval melanization prior to death was closely associated with the onset of high cysteine protease activity of ScathL in the hemolymph of fifth instars infected with AcMLF9.ScathL, but not with AcMLF9.ScathL.C146A, a recombinant baculovirus expressing a catalytic site mutant of ScathL. Fragmented fat body, ruptured gut and malpighian tubules, and melanized tracheae were observed in AcMLF9.ScathL-infected larvae. Phenoloxidase activity in hemolymph was unchanged, but the pool of prophenoloxidase was significantly reduced in virus-infected larvae and further reduced in AcMLF9.ScathL-infected larvae. The median lethal dose (LD(50)) for purified ScathL injected into fifth-instar H. virescens was 11.0 microg/larva. ScathL was also lethal to adult pea aphids, Acyrthosiphon pisum with a similar loss of integrity of the gut and fat body. Injection with purified ScathL.C146A or bovine trypsin at 20 microg/larva did not produce any effect in either insect. These results illustrate the potent insecticidal effects of ScathL cysteine protease activity and the potential for use of ScathL in development of insect resistant transgenic plants when combined with an appropriate delivery system.},
	keywords = {Animals, Baculoviridae, Basement Membrane/*drug effects, Biological Assay, Catechol Oxidase/blood, Cathepsins/genetics/*toxicity, Cysteine Endopeptidases/genetics/*toxicity, Diptera/chemistry, Enzyme Precursors/blood, Hemiptera/*drug effects, Insecticides/*toxicity, Larva/drug effects/metabolism, Lethal Dose 50, Moths/*drug effects/metabolism},
	author = {Li, H. and Tang, H. and Sivakumar, S. and Philip, J. and Harrison, R. L. and Gatehouse, J. A. and Bonning, B. C.}
}
