Palmitoylation of Intracellular Signaling Proteins: Regulation and Function

Smotrys, Jessica E.; Linder, Maurine E.

doi:10.1146/annurev.biochem.73.011303.073954

Cited by 543 publications

(510 citation statements)

References 174 publications

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“…Very recently, it has been shown that nSMase2 is S-palmitoylated in two Cys clusters; one cluster was located between the two hydrophobic segments, and the second one located in the middle of catalytic region of the protein [7]. Since S-palmitoylation is mostly restricted to the cytoplasmic side of the membranes [13], this posttranslational modification strongly supports the model presented in this study (proposed membrane topology and palmitoylation sites are shown in Fig. 6).…”

Section: Discussionsupporting

confidence: 88%

Analysis of membrane topology of neutral sphingomyelinase 2

Tani

Hannun

2007

FEBS Letters

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Neutral sphingomyelinase 2 (nSMase2), which has two hydrophobic segments at its NH 2 -terminus, plays an important role in ceramide-mediated cell regulation. Here, we investigated the membrane topology of nSMase2. When a double-tagged nSMase2 at both the NH 2 and COOH termini, was overexpressed in MCF-7 cells, the signals from both tags were detected in the inner leaflet of the plasma membrane. Furthermore, insertion of a tag into the internal sequence and green fluorescent protein-fused deletion mutants revealed that the entire catalytic region of the protein was located on the cytosolic face of the membranes and each hydrophobic segment is integrated into the membranes, but unlikely to span the entire membrane. These results indicate the presence of the enzyme in the inner leaflet of plasma membrane.

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Section: Discussionsupporting

confidence: 88%

Analysis of membrane topology of neutral sphingomyelinase 2

Tani

Hannun

2007

FEBS Letters

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“…Palmitoylation of proteins has several possible functions, depending on the protein under scrutiny. Much like protein prenylation in general, protein palmitoylation enables or contributes to membrane binding and possibly localization 52,53 . Additionally, palmitoylation may modulate protein-protein or protein-lipid interactions and enzyme activity.…”

Section: Palmitoylation Of Prenylated Proteinsmentioning

confidence: 99%

Therapeutic intervention based on protein prenylation and associated modifications

et al. 2006

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In eukaryotic cells, a specific set of proteins are modified by C-terminal attachment of 15-carbon farnesyl groups or 20-carbon geranylgeranyl groups that function both as anchors for fixing proteins to membranes and as molecular handles for facilitating binding of these lipidated proteins to other proteins. Additional modification of these prenylated proteins includes C-terminal proteolysis and methylation, and attachment of a 16-carbon palmitoyl group; these modifications augment membrane anchoring and alter the dynamics of movement of proteins between different cellular membrane compartments. The enzymes in the protein prenylation pathway have been isolated and characterized. Blocking protein prenylation is proving to be therapeutically useful for the treatment of certain cancers, infection by protozoan parasites and the rare genetic disease Hutchinson-Gilford progeria syndrome.Isoprenoids are lipids made of five-carbon blocks, and they constitute one of the main classes of natural products. A subset of isoprenoids called prenyl groups are found on a variety of biological substances, including proteins. Prenylated proteins and peptides are found in most (if not all) eukaryotes. They arise from the post-translational attachment of 15-carbon farnesyl or 20-carbon geranylgeranyl groups to the C-terminal segment of proteins. Protein prenyl groups not only are hydrophobic elements that bind proteins to membranes, but, at least in some cases, they also function as molecular handles that bind to hydrophobic grooves on the surface of soluble protein factors; these factors then remove the prenylated protein from the membrane in a regulated manner. NIH Public Access Author ManuscriptNat Chem Biol. Author manuscript; available in PMC 2010 June 28. Published in final edited form as:Nat Chem Biol. 2006 October ; 2(10): 518-528. doi:10.1038/nchembio818. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptProtein prenylation has been vigorously studied over the past ~15 years because it is found on several signaling proteins (including heterotrimeric G proteins) that connect cell surface receptors to intracellular effectors, and also on Ras proteins, one of the most common oncoproteins found in human tumors. Ras proteins serve as molecular switches at cellular membranes to control propagation of growth signals from cell surface receptors to nuclear transcription factors. Because Ras mutations are important in many human cancers, there has been extensive focus on Ras prenylation. Discovery of protein prenyl groups and structural varietiesThe first reports of prenylated proteins and peptides described the secreted pheromone peptides from jelly fungi 1,2 , whose structure resembles that of the well-known a-factor mating pheromone from baker's yeast (Saccharomyces cerevisiae), which contains a cysteine methylester farnesylated at the C terminus 3 . In the 1980s, studies on the timing of cholesterol biosynthesis with respect to the cell cycle in human cells led to the discovery that a compound deriv...

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“…Increasing numbers of proteins have been shown to be covalently modified by the 16 carbon saturated fatty acid palmitate [1][2][3]. Attachment of palmitate to proteins occurs primarily via thio-ester linkage to cysteine (S-palmitoylation).…”

Section: Introductionmentioning

confidence: 99%

“…The saturated nature of the palmitoyl chain promotes insertion of the fatty acylated protein into liquid ordered membrane domains [10]. Palmitoylation also influences intracellular protein trafficking and targeting to specific subcellular membranes, protein-protein interactions and protein activity [3,11].…”

Section: Introductionmentioning

confidence: 99%

Use of analogs and inhibitors to study the functional significance of protein palmitoylation

Resh

2006

Methods

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Covalent attachment of palmitate to proteins is a post-translational modification that exerts diverse effects on protein localization and function. The three key technical approaches required for an investigator to determine the role of palmitoylation of Your Favorite Palmitoylated Protein (YFPP) are methods to: 1) detect YFPP palmitoylation; 2) alter or inhibit palmitoylation of YFPP; 3) determine the functional significance of altered YFPP palmitoylation. Here, I describe experimental methods to address these three issues. Both radioactive (radiolabeling with 3 H palmitate or 125 I-IC16 palmitate) and non-radioactive (chemical labeling and mass spectrometry) methods to detect palmitoylated proteins are presented. Next, techniques to inhibit protein palmitoylation are described. These include site specific mutagenesis, and treatment of cells with inhibitors of protein palmitoylation, including 2-bromopalmitate, cerulenin, and tunicamycin. Alternative methods to replace palmitate with other fatty acids are also presented. Finally, general approaches to determining the effect of altered palmitoylation status on YFPP association with membranes and lipid rafts, as well as signal transduction, are described.

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Palmitoylation of Intracellular Signaling Proteins: Regulation and Function

Cited by 543 publications

References 174 publications

Analysis of membrane topology of neutral sphingomyelinase 2

Analysis of membrane topology of neutral sphingomyelinase 2

Therapeutic intervention based on protein prenylation and associated modifications

Use of analogs and inhibitors to study the functional significance of protein palmitoylation

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