Cysteine34 of the cytoplasmic tail of the cation-dependent mannose 6-phosphate receptor is reversibly palmitoylated and required for normal trafficking and lysosomal enzyme sorting.

Schweizer, Anja; Kornfeld, Stuart; Rohrer, Jack

doi:10.1083/jcb.132.4.577

Cited by 94 publications

(58 citation statements)

References 44 publications

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“…It was also noticed that mutation of either Cys&&) or Cys&'$ almost completely abolished palmitoylation, suggesting that acylation of both cysteines is synergistic. This contrasts with the CD-MPR, where the loss of palmitoylation of one cysteine was compensated by increased palmitoylation of the other [28].…”

Section: Discussionmentioning

confidence: 79%

“…Comparison of the half-lives of the palmitate moiety in other proteins with the half-lives of their respective polypeptide backbones indicates that there is large variation : by a factor of two for the transferrin receptor (t "/# $ 6 h [44]), a factor of 20 for the CD-MPR (t "/# $ 2 h [28]) and by a factor of 75 or more for p21 N V ras (t "/# $ 20 min [45]) and ankyrin (t "/# $ 50 min [39]). Two protein palmitoyl thioesterases that deacylate proteins have been identified, palmitoyl-protein thioesterase 1 (PPT1) and acyl-protein thioesterase 1 (APT1) [43].…”

Section: Discussionmentioning

confidence: 99%

“…Despite the presence of small amounts of LPC at the cell surface under steady-state conditions, abnormal cycling between the TGN and the plasma membrane might have a pronounced effect on the half-life of LPC. For instance, it has been shown that palmitoylation of the CD-MPR prevents the receptor from entering the lysosomes during its trafficking between the plasma membrane, endosomes and the TGN [28].…”

Section: Discussionmentioning

confidence: 99%

“…In contrast with the chemical stability of the acyl-amide bond in N-myristoylation [27], the chemically labile thioester bond allows regulated cycles of palmitoylation and depalmitoylation that may control a protein's subcellular localization. In the case of the cation-dependent mannose-6-phosphate receptor (CD-MPR), palmitoylation modulates sorting from endosomes to the TGN [28]. Palmitoylation of the transferrin receptor might reduce its endocytosis rate [29,30].…”

Section: Introductionmentioning

confidence: 99%

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Dynamic palmitoylation of lymphoma proprotein convertase prolongs its half-life, but is not essential for trans-Golgi network localization

LOO

TEUCHERT

Pauli

et al. 2000

Biochemical Journal

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Proprotein convertases are responsible for the endoproteolytic activation of proproteins in the secretory pathway. The most recently discovered member of this family, lymphoma proprotein convertase (LPC), is a type-I transmembrane protein. Previously, we have demonstrated that its cytoplasmic tail is palmitoylated. In this study, we have identified the two most proximal cysteine residues in the cytoplasmic tail as palmitoylation sites. Substitution of either cysteine residue by alanine interfered with palmitoylation of the other. Palmitoylation of LPC was found to be sensitive to the protein palmitoyltransferase inhibitor tunicamycin but not cerulenin. It was also insensitive to the drugs brefeldin A, monensin and cycloheximide, indicating that the modification occurs in a late exocytic or endocytic compartment. Turnover of palmitoylated LPC is significantly faster (t1/2 ≈ 50min) than that of the LPC polypeptide backbone (t1/2 ≈ 3h), suggesting that palmitoylation is reversible. Abrogation of palmitoylation reduced the half-life of the LPC protein, but did not affect steady-state localization of LPC in the trans-Golgi network. Finally, LPC could not be detected in detergent-resistant membrane rafts. Taken together, these results suggest that dynamic palmitoylation of LPC is important for stability, but does not function as a dominant trafficking signal.

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Section: Discussionmentioning

confidence: 79%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Dynamic palmitoylation of lymphoma proprotein convertase prolongs its half-life, but is not essential for trans-Golgi network localization

LOO

TEUCHERT

Pauli

et al. 2000

Biochemical Journal

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“…1 D and E). Mutations of palmitoylation sites often lead to the aberrant palmitoylation of neighboring cysteines (8,15). Therefore, we can only conclude that human LRP6 is palmitoy- lated on Cys-1394 and/or Cys-1399.…”

Section: Lrp6 Is Palmitoylated On a Juxtamembranous Cysteinementioning

confidence: 86%

Palmitoylation and ubiquitination regulate exit of the Wnt signaling protein LRP6 from the endoplasmic reticulum

Abrami

Kunz

Iacovache

et al. 2008

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

151

175

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Canonical Wnt signaling is initiated by binding of Wnt proteins to members of the Frizzled family and subsequent complex formation with lipoprotein receptor-related proteins 5/6 (LRP5/6). Here, we show that LRP6 is palmitoylated on a juxtamembranous cysteine and that palmitoylation is required for exit from the endoplasmic reticulum (ER). We propose that palmitoylation serves to tilt the long, 23-residue transmembrane domain of LRP6 with respect to the plane of membrane to prevent a hydrophobic mismatch and subsequent recognition by the ER quality control. In support of this model, a palmitoylation-deficient LRP6 mutant could be rescued from ER retention by deletion of two to four residues in the transmembrane domain. Importantly, we found that palmitoylation-deficient LRP6 was retained in the ER by a completely novel monoubiquitinationdependent ER retention mechanism. Mutation of a specific lysine indeed abolished ubiquitination of palmitoylation-deficient LRP6 and led to a rescue from ER retention. Finally, at the cell surface, we found that interplay between palmitoylation and ubiquitination was necessary for efficient Wnt signaling.he signaling of Wnt plays a critical role in a variety of developmental and adult processes in all metazoan and in diseases such as cancer (1). Canonical Wnt signaling via ␤-catenin is transduced by two receptor families: the Frizzled proteins and lipoprotein receptor-related proteins (LRPs) 5 and 6. Wnt binds to Frizzled, which then associates with LRP5/6. The cytoplasmic domains of LRP5/6, upon receptor activation by Wnt proteins, recruit the cytosolic scaffold protein axin to the membrane, displacing it from the so-called destruction complex (containing axin, glycogen synthase kinase 3␤, Wilms tumor suppressor, and ␤-catenin) involved in the degradation of ␤-catenin by the proteasome (for review see ref.2). This rescue of ␤-catenin allows it to translocate into the nucleus and interact with T cell factor (TCF)/lymphoid enhancer factor to activate transcription (3). As a result, Wnt activates multiple intracellular cascades, leading to cell differentiation, proliferation, migration, and polarity.LRP6 is a type I membrane protein with a large, 1,351-aa extracellular domain, containing multiple ␤-propeller and EGF domains, and a 220-aa-long cytoplasmic tail. As all type I transmembrane proteins, LRP6 is synthesized with an Nterminal signal sequence that targets it to the endoplasmic reticulum (ER). Folding of the LRP6 ␤-propeller domains in the lumen of the ER requires the help of an ER-resident chaperone called mesoderm development candidate 2 (Mesd) in mammals and Boca in Drosophila (4, 5). In the absence of Mesd, LRP6 is retained in the ER and thus fails to reach the cell surface.Human LRP6 contains six cysteine residues in its cytoplasmic tail, which are potential sites for palmitoylation (6). This lipid can serve to tether soluble proteins to membranes but is also found attached to the cytoplasmic domains of membrane proteins often close to the transmembrane region (7). The f...

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Analysis of Protein Transport to Lysosomes

2005

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Lysosomes are membrane-bound degradative organelles that are found in all higher eukaryotic cells. The limiting membranes of lysosomes contain a characteristic set of transmembrane glycoproteins that are transported to lysosomes by virtue of lysosomal targeting motifs found in the proteins' cytoplasmic tails (Bonifacino and Dell'angelica, 1999) The lumens of lysosomes contain an assortment of acidic hydrolases. These enzymes are synthesized in the endoplasmic reticulum (ER), where they become core-glycosylated. They are then transported into the Golgi complex, where they acquire a mannose-6-phosphate recognition tag in a two-step enzymatic process mediated by a phosphotransferase and an uncovering enzyme. The tag is then recognized by cation-dependent and cation-independent mannose-6-phoshate receptors in the trans-Golgi network (TGN). The resulting enzyme-receptor complexes are transported onward to endosomes, where the acidic pH induces dissociation of the enzymes from the receptors. The enzymes are then transported to lysosomes, while the receptors recycle back to the TGN to facilitate further rounds of transport. Failure of the receptors to recycle results in their misrouting to (and subsequent degradation in) lysosomes. The return of these receptors from endosomes to the TGN depends on specific amino acid residues (including specific cysteine residues that undergo reversible palmitoylation) in the cytoplasmic tails of receptor molecules. The final delivery of lysosomal enzymes to lysosomes has been proposed to occur by a fusion-fission process, referred to as "kiss and run," involving late endosomes and lysosomes (Luzio et al., 2003). BASIC PROTOCOL FRACTIONATION OF CELLS USING A SELF-FORMING PERCOLL DENSITY GRADIENT Sedimentation on self-forming Percoll gradients can be used to separate dense lysosomes from lower-density organelles such as endosomes, Golgi apparatus, plasma membrane, and ER. Separation is based on the sedimentation of particles to their isopycnic positioni.e., the position where the gradient density is equal to the density of the particle. The organelles are thus separated solely on the basis of differences in density, irrespective of their size. The gradient described in this protocol is a continuous gradient that forms

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Cysteine34 of the cytoplasmic tail of the cation-dependent mannose 6-phosphate receptor is reversibly palmitoylated and required for normal trafficking and lysosomal enzyme sorting.

Cited by 94 publications

References 44 publications

Dynamic palmitoylation of lymphoma proprotein convertase prolongs its half-life, but is not essential for trans-Golgi network localization

Dynamic palmitoylation of lymphoma proprotein convertase prolongs its half-life, but is not essential for trans-Golgi network localization

Palmitoylation and ubiquitination regulate exit of the Wnt signaling protein LRP6 from the endoplasmic reticulum

Analysis of Protein Transport to Lysosomes

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