| Literature DB >> 24365170 |
Ashapogu Venugopal1, Nadimpalli Siva Kumar2.
Abstract
A lysosomal cathepsin D (EC 3.4.23.5) was purified to homogeneity from the soft tissues of the fresh water mussel (Lamellidens corrianus) by pepstatin A affinity chromatography. The purified enzyme is a glycoprotein and migrates as a single protein species in native PAGE and shows a single band in SDS-PAGE corresponding to a molecular mass of ~43 kDa. Under both these conditions cathepsin D hydrolyzes hemoglobin as shown by zymogram analysis. The purified enzyme cross-reacts with an antiserum to purified starfish (Asterias rubens) cathepsin D. Additionally, the enzyme was recognized by the starfish lysosomal enzyme targeting receptors (mannose 6-phosphate receptors: MPR 300 and 46) in ligand blot analysis. The KM value of the purified enzyme with hemoglobin is 1.5mM. pH and temperature optimum for the enzyme are 3.5 and 60 °C respectively.Entities:
Keywords: 5-bromo-4-chloro-3′-indolyl phosphate; BCIP; Cathepsin D; ECL; Hb; IU; Invertebrate; Lamellidens corrianus; MPR; Mannose 6-phosphate receptor; NBT; PBST; TCA; Zymogram; enhanced chemiluminescence; hemoglobin; international unit; mannose 6-phosphate receptor; nitro blue tetrazolium chloride; phosphate-buffered saline containing Tween 20; trichloroacetic acid
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Year: 2013 PMID： 24365170 DOI： 10.1016/j.cbpb.2013.12.003
Source DB: PubMed Journal: Comp Biochem Physiol B Biochem Mol Biol ISSN： 1096-4959 Impact factor: 2.231