Purification and biochemical characterization of a lysosomal α-fucosidase from the deuterostomia Asterias rubens

被引：6

作者：

Visa, Merino ^{[1
]}

Hammer, Elke ^{[2
]}

Voelker, Uwe ^{[2
]}

Koliwer-Brandl, Hendrik ^{[3
]}

Kelm, Sorge ^{[3
]}

Nadimpalli, Siva Kumar ^{[1
]}

机构：

[1] Univ Hyderabad, Dept Biochem, Prot Biochem & Mol Biol Lab, Hyderabad 500046, Andhra Pradesh, India

[2] Ernst Moritz Arndt Univ Greifswald, Interfac Inst Genet & Funct Genom, D-17487 Greifswald, Germany

[3] Univ Bremen, Dept Biol & Chem, Ctr Biomol Interact Bremen, D-28359 Bremen, Germany

来源：

BIOCHIMIE | 2012年 / 94卷 / 05期

关键词：

alpha-Fucosidase; Invertebrate; LC-MS/MS; De novo sequencing; MPR300; RECEPTOR; TRANSPORT; PROTEINS; ENZYMES;

D O I：

10.1016/j.biochi.2012.02.007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In vertebrates, mannose 6-phosphate receptors [MPR300 (Mr 300 kDa) and MPR46 (Mr 46 kDa); are highly conserved transmembrane glycoproteins that mediate transport of lysosomal enzymes to lysosomes. Our studies have revealed the appearance of these putative receptors in invertebrates such as the molluscs and deuterostomes. Starfish tissue extracts contain several lysosomal enzyme activities and here we describe the affinity purification of alpha-fucosidase. The purified enzyme is a glycoprotein that exhibited a molecular mass of similar to 56 kDa in SOS-PAGE under reducing conditions. It has also cross-reacted with an antiserum to the mollusc enzyme suggesting antigenic similarities among the two invertebrate enzymes. LC-MS/MS analysis of the proteolytic peptides of the purified enzyme in combination with de novo sequencing allowed us to do partial amino acid sequence determination of the enzyme. These data suggest that this invertebrate enzyme is homologous to the known mammalian enzyme. The purified enzyme exhibited a mannose 6-phosphate dependent interaction with the immobilized starfish MPR300 protein. Our results demonstrate that the lysosomal enzyme targeting pathway is conserved even among the invertebrates. (C) 2012 Elsevier Masson SAS. All rights reserved.

引用

页码：1199 / 1205

页数：7

共 28 条

[1] Characterization of the mannose 6-phosphate receptor (Mr 300 kDa) protein dependent pathway of lysosomal enzyme targeting in Biomphalaria glabrata mollusc cells [J].