PDBsum entry 1mvv

Hydrolase

PDB id: 1mvv

Contents

Protein chain

313 a.a.

Ligands

M6P ×558

M6P-M6P ×10

Theoretical model

PDB id:

1mvv

Name:

Hydrolase

Title:

Theoretical structure of mouse cathepsin l with location of phosphorylatable o-6 atoms

Structure:

Cathepsin l. Chain: a. Ec: 3.4.22.15

Source:

Mus musculus. Mouse

Authors:

J.B.Warner,C.Thalhauser,K.Tao,G.G.Sahagian

Key ref:

J.B.Warner et al. (2002). Role of N-linked oligosaccharide flexibility in mannose phosphorylation of lysosomal enzyme cathepsin L. J Biol Chem, 277, 41897-41905. PubMed id: 12202476 DOI: 10.1074/jbc.M203097200

Date:

26-Sep-02 Release date: 31-Dec-02

Protein chain

P06797  (CATL1_MOUSE) -  Cathepsin L1

Seq: 334 a.a.

Struc: 313 a.a.

DOI no: 10.1074/jbc.M203097200

J Biol Chem 277:41897-41905 (2002)

PubMed id: 12202476

Role of N-linked oligosaccharide flexibility in mannose phosphorylation of lysosomal enzyme cathepsin L.

J.B.Warner, C.Thalhauser, K.Tao, G.G.Sahagian.

ABSTRACT

Mannose phosphorylation of N-linked oligosaccharides by UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase is a key step in the targeting of lysosomal enzymes in mammalian cells and tissues. The selectivity of this process is determined by lysine-based phosphorylation signals shared by lysosomal enzymes of diverse structure and function. By introducing new glycosylation sites at several locations on the surface of mouse procathepsin L and modeling oligosaccharide conformations for sites that are phosphorylated, it was shown that the inherent flexibility of N-linked oligosaccharides can account for the specificity of the transferase for oligosaccharides at different locations on the protein. By using this approach, the physical relationship between the lysine-based signal and the site of phosphorylation of mannose residues was determined. The analysis also revealed the existence of additional independent lysine-based phosphorylation signals on procathepsin L, which account for the low level of phosphorylation observed when the primary Lys-54/Lys-99 signal is ablated. Mutagenesis of residues that surround Lys-54 and Lys-99 and demonstration of mannose phosphorylation of a glycosylated derivative of green fluorescent protein provide strong evidence that the cathepsin L phosphorylation signal is a simple structure composed of as few as two well placed lysine residues.

Selected figure(s)

Figure 5.
Fig. 5. Location of the new glycosylation sites on cathepsin L. The wild-type glycosylation site (Asn-221) is shown in blue. The glycosylation sites that were added but were not significantly phosphorylated are shown in red. Glycosylation sites that are highly phosphorylated are shown in green. The -carbons of the critical lysines, Lys-54 and Lys-99, are shown in yellow.

Figure 6.
Fig. 6. Modeling of oligosaccharide clouds on cathepsin L. Modeled structures for oligosaccharide clouds of highly phosphorylated cathepsin L constructs are shown along with an overlay and overlap images. The overlap image is a superposition of the six individual structures. The overlap image shows the oligosaccharide region shared by the six individual structures. Oligosaccharide clouds were modeled as described under "Experimental Procedures." The -carbons of critical lysine residues Lys-54 and Lys-99 are displayed in white. WT, wild type.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2002, 277, 41897-41905) copyright 2002.

Figures were selected by an automated process.