@article {886,
	title = {Identification and purification of sulfotransferases for 20-hydroxysteroid from the larval fat body of a fleshfly, Sarcophaga peregrina},
	volume = {67},
	year = {Submitted},
	note = {08-10-200906-02-2012NormalKyoritsu College of Pharmacy, Minato-ku, Tokyo 105-8512, Japan.enghttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=12951514},
	pages = {1780 - 1785},
	abstract = {Sulfotransferase (ST) activity for 20-hydroxyecdysone (20E) was identified in a larval fat body lysate of the fleshfly, Sarcophaga peregrina, but not in the hemolymph. The activity was highly sensitive to 2,6-dichloro-4-nitrophenol (DCNP) (IC50=0.61 microM), a specific inhibitor of phenol ST (P-ST), but insensitive to triethylamine, a hydroxysteroid ST inhibitor. These results suggest that 20E-specific ST enzymes belong to the P-ST family, despite the fact that 20E is a hydroxysteroid. In addition to 20E ST activity, a relatively high level of 2-naphthol ST activity was detected in the fat body lysate. The ST activity for both substrates transiently decreased to the 50\% of maximal levels, 6 hrs after induction of pupation. The ST enzymes were separated on a DEAE-cellulose column. The 20E-ST enzymes were eluted around 50 mM KCl as two separate peaks of close proximity and the P-ST was eluted at 0.1 M KCl. The 20E ST enzymes were further purified using 3{\textquoteright}-phosphoadenosine 5{\textquoteright}-phosphate (PAP)-agarose affinity column chromatography. Both of the eluted active fractions demonstrated 43-kDa proteins on SDS-polyacrylamide gel. Photoaffinity labeling with [35S]-3{\textquoteright}-phosphoadenosine 5{\textquoteright}-phosphosulfate (PAPS) showed 43-kDa bands in the fat body lysate, as well as in the purified fractions. These results suggest that the 43-kDa proteins catalyze 20E sulfation within the fat body of S. peregrina.},
	keywords = {Adenosine Diphosphate/pharmacology, Animals, Chromatography, DEAE-Cellulose, derivatives/metabolism, development, Diptera/*enzymology/growth \&amp, Ecdysterone/antagonists \&amp, Enzyme Inhibitors/chemistry/pharmacology, Fat Body/chemistry/*enzymology, Hemolymph/metabolism, inhibitors/*chemistry/isolation \&amp, inhibitors/isolation \&amp, Larva/chemistry/enzymology, Naphthols/isolation \&amp, Nitrophenols/pharmacology, Phosphoadenosine Phosphosulfate/analogs \&amp, Photoaffinity Labels/chemistry/metabolism, Pupa/metabolism, purification/*metabolism, purification/metabolism, Sulfates/metabolism, Sulfotransferases/antagonists \&amp, Sulfur Radioisotopes},
	author = {Matsumoto, E. and Matsui, M. and Tamura, H. O.}
}
@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 {470,
	title = {p38 MAPK is a likely component of the signal transduction pathway triggering rapid cold hardening in the flesh fly Sarcophaga crassipalpis},
	volume = {210},
	year = {Submitted},
	note = {08-10-200906-02-2012NormalDepartment of Entomology, Ohio State University, 400 Aronoff Laboratory, 318 West 12th Avenue, Columbus, OH 43210, USA. fujiwara.6@osu.eduenghttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=17766307},
	pages = {3295 - 3300},
	abstract = {Rapid cold hardening (RCH) is an adaptation enabling insects to quickly respond to low temperature, but little is known about the molecular events that trigger this response. In this study of the flesh fly Sarcophaga crassipalpis, we explore a possible role for mitogen-activated protein kinases (MAPKs) in the low temperature signaling that elicits RCH. We report that p38 MAPK from S. crassipalpis, which shows high cDNA sequence homology to p38 MAPKs from other insects and mammals, is rapidly activated at temperatures around 0 degrees C, temperatures that are most effective for inducing RCH. By contrast, low temperature does not activate either extracellular signal-regulated kinase (ERK) or Jun N-terminal kinase (JNK). An increase in phospho-p38 MAPK was observed within 10 min following exposure to 0 degrees C and reached its maximum level in 2 h. When flies were transferred from 0 to 25 degrees C, the level of phospho-p38 MAPK decreased immediately and reached trace levels by 3 h. Nondiapausing flies were much more responsive to p38 MAPK activation than cold-hardy diapausing pupae. Thus, p38 MAPK activation and RCH both show the same narrow ranges of temperature sensitivity, temporal profiles of activation and decay, and developmental specificity. These correlations suggest that p38 MAPK plays a potential role in regulating the induction of RCH. The p38 MAPK response was not dependent upon the brain, as evidenced by high activation in isolated abdomens exposed to low temperature.},
	keywords = {*Cold Temperature, Acclimatization/*physiology, Amino Acid Sequence, Animals, Cloning, Molecular, development/*physiology, Diptera/*enzymology/growth \&amp, Enzyme Activation, p38 Mitogen-Activated Protein Kinases/*metabolism, Phosphorylation, Pupa, Signal Transduction/*physiology},
	author = {Fujiwara, Y. and Denlinger, D. L.}
}
