title: A digestive prolyl carboxypeptidase in Tenebrio molitor larvae.
abstract: Prolyl carboxypeptidase (PRCP) is a lysosomal proline specific serine peptidase that also plays a vital role in the regulation of physiological processes in mammals. In this report, we isolate and characterize the first PRCP in an insect. PRCP was purified from the anterior midgut of larvae of a stored product pest, Tenebrio molitor, using a three-step chromatography strategy, and it was determined that the purified enzyme was a dimer. The cDNA of PRCP was cloned and sequenced, and the predicted protein was identical to the proteomic sequences of the purified enzyme. The substrate specificity and kinetic parameters of the enzyme were determined. The T. molitor PRCP participates in the hydrolysis of the insect's major dietary proteins, gliadins, and is the first PRCP to be ascribed a digestive function. Our collective data suggest that the evolutionary enrichment of the digestive peptidase complex in insects with an area of acidic to neutral pH in the midgut is a result of the incorporation of lysosomal peptidases, including PRCP.
authors.affiliation: Chemical Faculty, Moscow State University, Moscow 119991, Russia.
authors.forename: Irina A
authors.initials: IA
authors.lastname: Goptar
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Shagin
authors.forename: Irina A
authors.initials: IA
authors.lastname: Shagina
authors.forename: Elena S
authors.initials: ES
authors.lastname: Mudrik
authors.forename: Yulia A
authors.initials: YA
authors.lastname: Smirnova
authors.forename: Dmitry P
authors.initials: DP
authors.lastname: Zhuzhikov
authors.forename: Mikhail A
authors.initials: MA
authors.lastname: Belozersky
authors.forename: Yakov E
authors.initials: YE
authors.lastname: Dunaevsky
authors.forename: Brenda
authors.initials: B
authors.lastname: Oppert
authors.forename: Irina Yu
authors.initials: IY
authors.lastname: Filippova
authors.forename: Elena N
authors.initials: EN
authors.lastname: Elpidina
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002268:Carboxypeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004064:Digestive System / Q000201:enzymology*; D006868:Hydrolysis; D007814:Larva / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D064828:Prolyl Hydroxylases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D013379:Substrate Specificity; D013712:Tenebrio / Q000201:enzymology* / Q000235:genetics
chemical_list: D064828:Prolyl Hydroxylases; D002268:Carboxypeptidases