Several cooperating binding sites mediate the interaction of a lysosomal enzyme with phosphotransferase

Tikkanen, Ritva; Peltola, Matti; Oinonen, Carita; Rouvinen, Juha; Peltonen, Leena

doi:10.1093/emboj/16.22.6684

Cited by 48 publications

(59 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The distance of ϳ34 Å between ␣-carbon atoms of critical lysine residues appears to be a general feature of the phosphorylation signal. This distance was demonstrated in our earlier study (17) for cathepsin D as well as cathepsin L (14) and is consistent with results obtained for two other proteins, DNase I (15, 29) and aspartylglucosaminidase (16). In the case of aspartylglucosaminidase, mutation of two lysine residues, Lys-183 in the ␣ subunit and Lys-214 in the ␤ subunit, was found to inhibit mannose phosphorylation by 96%.…”

Section: Discussionsupporting

confidence: 89%

“…On the basis of these results, a relatively simple model involving lysine residues was proposed as a general phosphorylation signal for lysosomal proteins. Similar involvement of lysine residues (15,16) has since been demonstrated for other proteins, and it is now widely accepted that lysine residues are the primary determinants for mannose phosphorylation of a wide range of proteins by the transferase.…”

supporting

confidence: 58%

“…for lysine in some contexts (15)(16)(17). Other residues in the vicinity of critical lysine residues may affect the accessibility or properties of the lysine residues.…”

Section: Discussionmentioning

confidence: 99%

“…Other residues in the vicinity of critical lysine residues may affect the accessibility or properties of the lysine residues. Effects such as this may explain the apparent involvement of tyrosine residues in phosphorylation of aspartylglucosaminidase and DNase I (16,29). In theory, any residue in contact with the transferase when it is bound to the protein could affect the rate and efficiency of phosphorylation.…”

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

Role of N-Linked Oligosaccharide Flexibility in Mannose Phosphorylation of Lysosomal Enzyme Cathepsin L

Warner

Thalhauser

Tao

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

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.The mammalian lysosomal protein targeting system has the capability of recognizing and modifying lysosomal hydrolases and growth factors from a wide range of protein families with high specificity. The molecular basis for this selectivity is due to the activity of the UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase), which phosphorylates N-linked oligosaccharides of these proteins by the addition of GlcNAc-1-P (1-5). This modification begins after lysosomal proteins are exported from the endoplasmic reticulum and is followed by the removal of the terminal GlcNAc moieties from the adducts. In the Golgi apparatus the phosphorylated proteins are bound to mannose 6-phosphate receptors, which mediate the delivery of the proteins to lysosomes.GlcNAc-1-phosphotransferase has been purified as a 540-kDa complex composed of disulfide-linked homodimers of ␣ and ␤ subunits and two identical, noncovalently associated ␥ subunits (6). The ␣ subunit was shown to have nucleotide sugar binding activity, and on the basis of previous genetic data, it was known that the catalytic and protein recognition activities are likely to be located on separate subunits (7). This has since been verified by analysis of the transferase in cells from patients with mucolipidosis IIIC or variant pseudo-Hurler polydystrophy. GlcNAc-1-phosphotransferase from these patients is defective in the ␥ subunit, which prevents phosphorylation of lysosomal enzymes, yet transferase activity on synthetic substrates is retained (8).Although the molecular basis for recognition of lysosomal hydrolases by the transferase has been studied qu...

show abstract

Section: Discussionsupporting

confidence: 89%

supporting

confidence: 58%

“…for lysine in some contexts (15)(16)(17). Other residues in the vicinity of critical lysine residues may affect the accessibility or properties of the lysine residues.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Role of N-Linked Oligosaccharide Flexibility in Mannose Phosphorylation of Lysosomal Enzyme Cathepsin L

Warner

Thalhauser

Tao

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…A number of studies from our laboratory and others have shown that the protein recognition domain of acid hydrolases is a complex conformation-dependent determinant that includes a number of critical lysine residues (16)(17)(18)(19)(20)(21)(22)(23). On this basis, it seems likely that the interaction of an acid hydrolase with GlcNAc-1-phosphotransferase would involve multiple contacts between the surfaces of the two proteins.…”

mentioning

confidence: 99%

The DMAP interaction domain of UDP-GlcNAc:lysosomal enzyme N -acetylglucosamine-1-phosphotransferase is a substrate recognition module

Qian

Flanagan-Steet

Meel

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase) is an α 2 β 2 γ 2 heterohexamer that mediates the initial step in the formation of the mannose 6-phosphate recognition signal on lysosomal acid hydrolases. We previously reported that the specificity of the reaction is determined by the ability of the α/β subunits to recognize a conformationdependent protein determinant present on the acid hydrolases. We now present evidence that the DNA methyltransferase-associated protein (DMAP) interaction domain of the α subunit functions in this recognition process. First, GST-DMAP pulled down several acid hydrolases, but not nonlysosomal glycoproteins. Second, recombinant GlcNAc-1-phosphotransferase containing a missense mutation in the DMAP interaction domain (Lys732Asn) identified in a patient with mucolipidosis II exhibited full activity toward the simple sugar α-methyl D-mannoside but impaired phosphorylation of acid hydrolases. Finally, unlike the WT enzyme, expression of the K732N mutant in a zebrafish model of mucolipidosis II failed to correct the phenotypic abnormalities. These results indicate that the DMAP interaction domain of the α subunit functions in the selective recognition of acid hydrolase substrates and provides an explanation for the impaired phosphorylation of acid hydrolases in a patient with mucolipidosis II.

show abstract

Identification of Motifs in Protein Sequences

Sonnhammer

Wolfsberg

1998

CP Cell Biology

View full text Add to dashboard Cite

This brief appendix serves as a guide for the analysis of functional motifs in proteins. Several database search engines that can be accessed via the World Wide Web are described. Such computerized searches have become the preferred method to scan large sequence and motif databases, as the searches are efficient and the databases are updated frequently. A short list of sorting signals is also included, since these motifs often cannot be predicted reliably by a computer search.

show abstract

Several cooperating binding sites mediate the interaction of a lysosomal enzyme with phosphotransferase

Cited by 48 publications

References 35 publications

Role of N-Linked Oligosaccharide Flexibility in Mannose Phosphorylation of Lysosomal Enzyme Cathepsin L

Role of N-Linked Oligosaccharide Flexibility in Mannose Phosphorylation of Lysosomal Enzyme Cathepsin L

The DMAP interaction domain of UDP-GlcNAc:lysosomal enzyme N -acetylglucosamine-1-phosphotransferase is a substrate recognition module

Identification of Motifs in Protein Sequences

Contact Info

Product

Resources

About