Human DHHC proteins: A spotlight on the hidden player of palmitoylation

doi:10.1016/j.ejcb.2011.09.013

European Journal of Cell Biology

Volume 91, Issue 2, February 2012, Pages 107-117

https://doi.org/10.1016/j.ejcb.2011.09.013 Get rights and content

Abstract

Palmitoylation is one of the most common posttranslational lipid modifications of proteins and we now know quite a lot about it. However, the state of knowledge about the enzymes that catalyze this process is clearly insufficient. This review is focused on 23 human DHHC genes and their products – protein palmitoyltransferases. Here we describe mainly the structure and function of these proteins, but also, to a lesser degree, what the substrates of the enzymes are and whether they are related to various diseases. The main aim of this review was to catalogue existing information concerning the human DHHC family of genes/proteins, making them and their functions easier to understand.

Section snippets

Palmitoylation at a glance

Palmitoylation was first described about 40 years ago and it is one of the most frequent posttranslational modifications of proteins, defined as the addition of saturated 16-carbon palmitic acid to specific cysteine residues. Depending on the type of palmitoylation, it occurs through an N-amide bond (N-palmitoylation) or through the formation of a labile thioester bond (S-palmitoylation). This review is focused only on S-palmitoylation. S-palmitoylation is a unique, reversible modification with

Human palmitoylating enzymes (PATs)

Palmitoylation is catalyzed by a family of enzymes that contain a DHHC (Asp-His-His-Cys) cysteine-rich domain which is directly involved in the palmitoyl transfer reaction (it is the apparent catalytic center of the enzyme) (Mitchell et al., 2006). The DHHC cysteine-rich domain is a highly conserved motif that can be found in multiple eukaryotic proteins (Ohno et al., 2006). This domain includes a region with the form of a Cys4 zinc-finger-like metal ion binding site followed by a complex

Human ZDHHC genes and proteins

Summarized information about the size of each gene, its location in the human genome and transcription products and also intracellular localization of proteins is presented in Table 1a, Table 1b. Most of the DHHC proteins are localized to the Golgi apparatus and to the endoplasmic reticulum (ER). However, some of them have been localized to the plasma membrane (Greaves et al., 2011, Ohno et al., 2006, Planey et al., 2009). The size of ZDHHC genes varies between 3.26 kb (ZDHHC12) and 296.68 kb (

ZDHHC1 and DHHC1

Very little is known about the human ZDHHC1 gene. According to the NCBI database (http://www.ncbi.nlm.nih.gov/AceView), the protein has probable catalytic activity as a palmitoyltransferase, but there are no known substrates for this enzyme. This gene is expressed at a high level in many human tissues, particularly in brain, lung and uterus (Table 1b). DHHC1 protein is expected to have molecular functions in the binding of DNA, proteins and metal ions (in particular zinc ions) (Ohno et al., 2006

ZDHHC2 and DHHC2

The human ZDHHC2 gene is expressed at the highest level in brain, lung, kidney and pancreas (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a). Moreover, this gene, originally named REAM (reduced expression associated with metastasis protein), maps to a region of chromosome 8 that is frequently deleted in many types of cancer, such as non-small cell lung cancers (Emi et al., 1993, Fujiwara et al., 1994), hepatocellular carcinoma (Bova et al., 1993, Fujiwara et al., 1994), colorectal cancer (Bova

ZDHHC3 and DHHC3

The human ZDHHC3 gene is expressed at a very high level in particular in liver, brain, spleen and lung (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a). DHHC3 is one of the Golgi-localized palmitoyltransferases (only a minimal overlap with the ER–Golgi intermediate compartment is observed), which was identified as an enzyme that can palmitoylate cysteine-string protein (CSP) and lead to its stable membrane binding (Greaves et al., 2008).

Furthermore, DHHC3 (also named GODZ) functions as a

ZDHHC4 and DHHC4

Information concerning the ZDHHC4 gene is rather poor (http://www.ncbi.nlm.nih.gov/AceView). All that is known nowadays is that the ZDHHC4 gene is expressed at a very high level mainly in brain and lung (Table 1b). Functionally, DHHC4 protein is expected to have acyltransferase activity, electron carrier activity (cytochrome-c oxidase activity) and metal ion binding capacity (particularly zinc and iron ions).

There is also evidence indicating that the mRNA level for DHHC4 protein is higher in

ZDHHC5 and DHHC5

Very little is known about the human ZDHHC5 gene. This gene is expressed at a high level mainly in prostate, testis and lung, but also in many other human tissues (Table 1b). According to the NCBI database (http://www.ncbi.nlm.nih.gov/AceView), the DHHC5 protein has acyltransferase activity and the capacity to bind metal ions. It is important to add that the in vivo function of the DHHC5 protein is still unknown, but there are data suggesting that DHHC5 (similar to DHHC6 and 8) is S-acylated on

ZDHHC6 and DHHC6

Expression of the human ZDHHC6 gene occurs at high levels, particularly in brain, uterus and kidney (Table 1b). DHHC6, like the other DHHC proteins, is expected to have acyltransferase activity and metal (zinc) ion binding capacity. As mentioned above, this protein also appears to be S-acylated on three cysteine residues within a CCX_7–13C(S/T) motif downstream of a conserved DHHC domain (Chen et al., 2004; http://www.ncbi.nlm.nih.gov/AceView; Yang et al., 2010).

ZDHHC7 and DHHC7

The ZDHHC7 gene is expressed at a high level mainly in lung, colon, brain and liver (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a), and the protein is the closest homolog of DHHC3 (90% identity within the DHHC domain). Like DHHC3 it is also a Golgi-localized palmitoyltransferase and specifically palmitoylates Gα protein in this compartment, which is necessary for the continuous Gα_q shuttling between the Golgi apparatus and the plasma membrane (Fang et al., 2006, Mill et al., 2009).

Moreover,

ZDHHC8 and DHHC8

ZDHHC8 gene expression occurs at a high level not only in the brain, but also in many other human tissues, for example in the eye and lung (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a). The ZDHHC8 gene probably encodes palmitoyltransferase, which is required for protein palmitoylation, regulating trafficking and signaling pathways that are essential for brain development and neurotransmitter receptors (el-Husseini Ael and Bredt, 2002, Otani et al., 2005).

The ZDHHC8 gene is located in the 1.5

ZDHHC9 and DHHC9

The ZDHHC9 gene is expressed at a high level, especially in the brain and prostate (http://www.ncbi.nlm.nih.gov/AceView; Raymond et al., 2007, Swarthout et al., 2005) (Table 1a).

For palmitoyltransferase activity and protein stability of DHHC9, its complex with the Golgi-localized membrane protein GCP16 is required (Mansilla et al., 2007, Swarthout et al., 2005). The DHHC9–GCP16 complex catalyzes the posttranslational modification of H- and N-Ras proteins. In the absence of GCP16 protein DHHC9

ZDHHC11 and DHHC11

The ZDHHC11 gene is well expressed mainly in placenta, testis and brain (Table 1b) and maps to the 5p15.33 region of chromosome 5. Gain of this region, including the ZDHHC11 gene, is a predictor for disease progression in bladder cancer, which may become a potential marker identifying high-risk patients with disease progression in this type of cancer (Planey et al., 2009, Yamamoto et al., 2007). Moreover, it was found that gain at chromosomal region 5p15.33, where the ZDHHC11 gene is located,

ZDHHC12 and DHHC12

The ZDHHC12 gene is expressed at a high level, especially in the stomach (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a). It was found that palmitoyltransferase DHHC12, also named AID/DHHC12 protein (APP interacting DHHC protein), serves to tether amyloid precursor protein (APP) in the Golgi apparatus and suppresses the generation of APP-containing vesicles, which in turn suppresses the trafficking of APP-containing vesicles into the late secretory pathway. This attenuated generation and

ZDHHC13 and DHHC13

The ZDHHC13 gene is expressed at a high level mainly in the uterus, but also in many other human tissues, for example in the stomach, placenta and testis (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a).

Palmitoyltransferase DHHC13, also named the HIP14L protein (HIP14-like protein), is one of two huntingtin-interacting proteins. It is the closest homolog of HIP14 (ZDHHC17), demonstrating 48% identity and 69% similarity (Goytain et al., 2008, Huang et al., 2009, Singaraja et al., 2002). Like the

ZDHHC14 and DHHC14

Expression of the ZDHHC14 gene occurs at a high level particularly in the brain and eye (http://www.ncbi.nlm.nih.gov/AceView) (Table 1b), but also in myeloid and lymphoid cells (in particular in B lymphocytes). The ZDHHC14 gene has been located to chromosome 6 in the 6q25.3 region. Gain in this region was detected in post-transplant lymphoproliferative disorders (PTLD) as well as in diffuse large B-cell lymphomas. Thus, the ZDHHC14 gene appears to be a putative oncogene in lymphomas, possibly

ZDHHC15 and DHHC15

The ZDHHC15 gene is well expressed mainly in the brain and trachea (Table 1b). It was found that the loss of ZDHHC15 expression is probably connected with X-linked mental retardation (XLMR) (Fang et al., 2006; http://www.ncbi.nlm.nih.gov/AceView; Mansouri et al., 2005).

DHHC15 protein has been localized mainly to the Golgi apparatus and is one of the DHHC enzymes that palmitoylate CSP and lead to its stable attachment to the membrane (Greaves et al., 2008). Other known substrates for this enzyme

ZDHHC16 and DHHC16

ZDHHC16 gene expression occurs at a high level mainly in the brain, placenta and lung (http://www.ncbi.nlm.nih.gov/AceView) (Table 1b). DHHC16 protein is expected to have palmitoyltransferase activity and the capacity to bind metal ions (especially zinc ions). It was discovered that DHHC16 interacts with non-receptor tyrosine kinase implicated in DNA damage-induced cell death and in growth factor receptor signaling named c-Abl and that this protein may be involved in apoptosis regulation (//www.phosphosite.org/proteinAction.do%3Fide=3067387%26showAllSites=true

ZDHHC17 and DHHC17

The ZDHHC17 gene is expressed at a high level particularly in the brain, but also in many other human tissues, for example in the uterus and eye (http://www.ncbi.nlm.nih.gov/AceView; Singaraja et al., 2002) (Table 1a). DHHC17, also known as HIP14, is a palmitoyltransferase which palmitoylates a number of neuronal proteins, including SNAP25 (Greaves et al., 2009), PSD-95, GAD65, synaptotagmin I and huntingtin (Fang et al., 2006, Gao et al., 2009, Huang et al., 2004, Huang et al., 2009, Stowers

ZDHHC18 and DHHC18

The ZDHHC18 gene is expressed at a very high level mainly in the lung, testis and brain (Table 1b). According to the NCBI database, DHHC18 protein has the capacity to bind metal ions and acyltransferase activity, but there are no known substrates for this enzyme (http://www.ncbi.nlm.nih.gov/AceView).

ZDHHC19 and DHHC19

The ZDHHC19 gene is well expressed. Expression of this gene occurs mainly in the testis, but also in the brain, medulla and placenta (http://www.ncbi.nlm.nih.gov/AceView) (Table 1b). It was found that DHHC19 is a putative palmitoyltransferase for R-Ras proteins. Palmitoylation of these proteins is necessary for their association with membranes as well as with lipid rafts (Baumgart et al., 2010).

ZDHHC20 and DHHC20

The ZDHHC20 gene is expressed at a high level in placenta, but also in many other human tissues (http://www.ncbi.nlm.nih.gov/AceView) (Table 1b). Recently it was demonstrated that the ZDHHC20 gene is overexpressed in several types of human tumors, including ovarian, breast and prostate. Moreover, it was shown that DHHC20 protein is human N-terminal-myristoyl-directed palmitoyltransferase, involved in cellular transformation, which may play a role in cancer (Draper and Smith, 2010).

ZDHHC21 and DHHC21

The ZDHHC21 gene is expressed at a high level mainly in the brain and testis (http://www.ncbi.nlm.nih.gov/AceView) (Table 1a). It shows palmitoyltransferase activity toward Fyn kinase (Mill et al., 2009). It is also one of the endothelial NO synthase PATs, which was found to be involved in the release of nitric oxide stimulated by both ionomycin and ATP. It is suggested that DHHC21 functions downstream of calcium and ATP (Fernandez-Hernando et al., 2006, Mill et al., 2009).

It is important to

ZDHHC22 and DHHC22

Information about the ZDHHC22 gene is rather scant. All that is known is that it is well expressed especially in the brain, eye and lung (Table 1b). The in vivo ZDHHC22 gene product remains unknown, but DHHC22 protein is expected to have catalytic activity as a palmitoyltransferase and it is capable of binding metal ions (mainly zinc ions) (http://www.ncbi.nlm.nih.gov/AceView).

ZDHHC23 and DHHC23

Very little is known about the human ZDHHC23 gene. According to the NCBI database (http://www.ncbi.nlm.nih.gov/AceView) it is expressed at a moderately high level, especially in the testis, brain, colon and kidney (Table 1b). DHHC23 protein is expected to have molecular functions such as binding metal (zinc) ions and acyltransferase activity. There is also one probable protein interactor for DHHC23, namely neuronal nitric oxide synthase 1 (NOS1) (nNOS), which was previously demonstrated in a

ZDHHC24 and DHHC24

The ZDHHC24 gene is also barely examined. As yet it is only known that it is well expressed mainly in the brain and pancreas, but also in the prostate and stomach (Table 1b) (http://www.ncbi.nlm.nih.gov/AceView).

The presence of ZDHHC gene transcripts in human blood cells and blood cell lines

According to the UniGene database, ZDHHC gene transcripts are present in many types of human blood cells and blood cell lines (http://www.ncbi.nlm.nih.gov/UniGene/browse2cgi?TAXID=9606) (Table 2). In some types of cells only one palmitoyltransferase is found (e.g. precursor erythroblasts, CD71+ progenitor cells or the HL60 cell line) and in others there are many ZDHHC gene transcripts (e.g. leukocytes, natural killer cells or the Jurkat T cell line).

Conclusions

As presented above, many DHHC proteins exist in the human/mammalian organism, where they differ in their intracellular localization as well as in tissue-specific expression. It was also found that ZDHHC genes are connected to certain pathologies; therefore they might be potential medical biomarkers or even drug targets. Undoubtedly, many questions remain unsolved and much work is still needed to elucidate the specific function(s) of each DHHC protein.

Acknowledgements

This work was supported by Grant No. N.N.301 1611435 from the Polish Ministry of Science and Higher Education. M.W. received a European Social Fund grant from the Office of the Marshal of the Lubuskie Voivodeship.

References (101)

F. Baumgart et al.
Palmitoylation of R-Ras by human DHHC19, a palmitoyl transferase with a CaaX box
Biochim. Biophys. Acta
(2010)
M.J. Bijlmakers et al.
The on-off story of protein palmitoylation
Trends Cell Biol.
(2003)
J.A. Duncan et al.
Autoacylation of G protein alpha subunits
J. Biol. Chem.
(1996)
J.T. Dunphy et al.
Differential effects of acyl-CoA binding protein on enzymatic and non-enzymatic thioacylation of protein and peptide substrates
Biochim. Biophys. Acta
(2000)
M. Fukata et al.
Identification of PSD-95 palmitoylating enzymes
Neuron
(2004)
Y. Fukata et al.
Systematic screening for palmitoyl transferase activity of the DHHC protein family in mammalian cells
Methods
(2006)
B. Glaser et al.
No association between the putative functional ZDHHC8 single nucleotide polymorphism rs175174 and schizophrenia in large European samples
Biol. Psychiatry
(2005)
A. Goytain et al.
Huntingtin-interacting proteins, HIP14 and HIP14L, mediate dual functions, palmitoyl acyltransferase and Mg²⁺ transport
J. Biol. Chem.
(2008)
J. Greaves et al.
DHHC palmitoyl transferases: substrate interactions and (patho)physiology
Trends Biochem. Sci.
(2011)
J. Greaves et al.
Palmitoylation of the SNAP25 protein family: specificity and regulation by DHHC palmitoyl transferases
J. Biol. Chem.
(2010)

J. Greaves et al.

Palmitoylation and membrane interactions of the neuroprotective chaperone cysteine-string protein

J. Biol. Chem.

(2008)

R.M. Hines et al.

Golgi-specific DHHC zinc finger protein GODZ mediates membrane Ca²⁺ transport

J. Biol. Chem.

(2010)

K. Huang et al.

Huntingtin-interacting protein HIP14 is a palmitoyl transferase involved in palmitoylation and trafficking of multiple neuronal proteins

Neuron

(2004)

B.C. Jennings et al.

2-Bromopalmitate and 2-(2-hydroxy-5-nitro-benzylidene)-benzo[b]thiophen-3-one inhibit DHHC-mediated palmitoylation in vitro

J. Lipid Res.

(2009)

J.U. Kang et al.

Gain at chromosomal region 5p15.33, containing TERT, is the most frequent genetic event in early stages of non-small cell lung cancer

Cancer Genet. Cytogenet.

(2008)

M. Karayiorgou et al.

The molecular genetics of the 22q11-associated schizophrenia

Brain Res. Mol. Brain Res.

(2004)

B. Li et al.

Aph2, a protein with a zf-DHHC motif, interacts with c-Abl and has pro-apoptotic activity

J. Biol. Chem.

(2002)

S. Lobo et al.

Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae

J. Biol. Chem.

(2002)

T.M. Maynard et al.

Mitochondrial localization and function of a subset of 22q11 deletion syndrome candidate genes

Mol. Cell. Neurosci.

(2008)

D.A. Mitchell et al.

Protein palmitoylation by a family of DHHC protein S-acyltransferases

J. Lipid Res.

(2006)

C. Montell

Mg²⁺ homeostasis: the Mg²⁺ nificent TRPM chanzymes

Curr. Biol.

(2003)

Y. Ohno et al.

Intracellular localization and tissue-specific distribution of human and yeast DHHC cysteine-rich domain-containing proteins

Biochim. Biophys. Acta

(2006)

K. Otani et al.

The ZDHHC8 gene did not associate with bipolar disorder or schizophrenia

Neurosci. Lett.

(2005)

F.L. Raymond et al.

Mutations in ZDHHC9, which encodes a palmitoyltransferase of NRAS and HRAS, cause X-linked mental retardation associated with a Marfanoid habitus

Am. J. Hum. Genet.

(2007)

M.D. Resh

Regulation of cellular signalling by fatty acid acylation and prenylation of signal transduction proteins

Cell. Signal.

(1996)

M.D. Resh

Fatty acylation of proteins: new insights into membrane targeting of myristoylated and palmitoylated proteins

Biochim. Biophys. Acta

(1999)

M.D. Resh

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

Methods

(2006)

S. Saito et al.

No association was found between a functional SNP in ZDHHC8 and schizophrenia in a Japanese case–control population

Neurosci. Lett.

(2005)

F. Saitoh et al.

NIDD, a novel DHHC-containing protein, targets neuronal nitric-oxide synthase (nNOS) to the synaptic membrane through a PDZ-dependent interaction and regulates nNOS activity

J. Biol. Chem.

(2004)

H. Sudo et al.

A loss of function screen identifies nine new radiation susceptibility genes

Biochem. Biophys. Res. Commun.

(2007)

J.T. Swarthout et al.

DHHC9 and GCP16 constitute a human protein fatty acyltransferase with specificity for H- and N-Ras

J. Biol. Chem.

(2005)

A. Undale et al.

Circulating osteogenic cells: characterization and relationship to rates of bone loss in postmenopausal women

Bone

(2010)

L.A. Verkruyse et al.

Lysosomal targeting of palmitoyl-protein thioesterase

J. Biol. Chem.

(1996)

W. Yang et al.

Proteome scale characterization of human S-acylated proteins in lipid raft-enriched and non-raft membranes

Mol. Cell. Proteomics

(2010)

F. Zhang et al.

Molecular cloning and characterization of human Aph2 gene, involved in AP-1 regulation by interaction with JAB1

Biochim. Biophys. Acta

(2006)

K. Anami et al.

Search for transmembrane protein in gastric cancer by the Escherichia coli ampicillin secretion trap: expression of DSC2 in gastric cancer with intestinal phenotype

J. Pathol.

(2010)

R. Anbazhagan et al.

Allelic loss of chromosomal arm 8p in breast cancer progression

Am. J. Pathol.

(1998)

P. Arstikaitis et al.

Paralemmin-1, a modulator of filopodia induction is required for spine maturation

Mol. Biol. Cell

(2008)

S. Baekkeskov et al.

Palmitoylation cycles and regulation of protein function (Review)

Mol. Membr. Biol.

(2009)

G.S. Bova et al.

Homozygous deletion and frequent allelic loss of chromosome 8p22 loci in human prostate cancer

Cancer Res.

(1993)

W.Y. Chen et al.

Case–control study and transmission disequilibrium test provide consistent evidence for association between schizophrenia and genetic variation in the 22q11 gene ZDHHC8

Hum. Mol. Genet.

(2004)

Z.W. Chen et al.

GABAA receptor associated proteins: a key factor regulating GABAA receptor function

J. Neurochem.

(2007)

E. Conibear et al.

Palmitoylation and depalmitoylation dynamics at a glance

J. Cell Sci.

(2010)

M.B. Dalva

Neuronal activity moves protein palmitoylation into the synapse

J. Cell Biol.

(2009)

F.J. Dekker et al.

Small-molecule inhibition of APT1 affects Ras localization and signaling

Nat. Chem. Biol.

(2010)

L.E. Dietrich et al.

On the mechanism of protein palmitoylation

EMBO Rep.

(2004)

J.M. Draper et al.

Palmitoyl acyltransferase assays and inhibitors (Review)

Mol. Membr. Biol.

(2009)

J.M. Draper et al.

DHHC20: a human palmitoyl acyltransferase that causes cellular transformation

Mol. Membr. Biol.

(2010)

C.E. Ducker et al.

Discovery and characterization of inhibitors of human palmitoyl acyltransferases

Mol. Cancer Ther.

(2006)

C.E. Ducker et al.

Huntingtin interacting protein 14 is an oncogenic human protein: palmitoyl acyltransferase

Oncogene

(2004)

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Modification of Proteins by Metabolites in Immunity
2021, Immunity
Immunometabolism has emerged as a key focus for immunologists, with metabolic change in immune cells becoming as important a determinant for specific immune effector responses as discrete signaling pathways. A key output for these changes involves post-translational modification (PTM) of proteins by metabolites. Products of glycolysis and Krebs cycle pathways can mediate these events, as can lipids, amino acids, and polyamines. A rich and diverse set of PTMs in macrophages and T cells has been uncovered, altering phenotype and modulating immunity and inflammation in different contexts. We review the recent findings in this area and speculate whether they could be of use in the effort to develop therapeutics for immune-related diseases.

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ReviewHuman DHHC proteins: A spotlight on the hidden player of palmitoylation

Abstract

Section snippets

Palmitoylation at a glance

Human palmitoylating enzymes (PATs)

Human ZDHHC genes and proteins

ZDHHC1 and DHHC1

ZDHHC2 and DHHC2

ZDHHC3 and DHHC3

ZDHHC4 and DHHC4

ZDHHC5 and DHHC5

ZDHHC6 and DHHC6

ZDHHC7 and DHHC7

ZDHHC8 and DHHC8

ZDHHC9 and DHHC9

ZDHHC11 and DHHC11

ZDHHC12 and DHHC12

ZDHHC13 and DHHC13

ZDHHC14 and DHHC14

ZDHHC15 and DHHC15

ZDHHC16 and DHHC16

ZDHHC17 and DHHC17

ZDHHC18 and DHHC18

ZDHHC19 and DHHC19

ZDHHC20 and DHHC20

ZDHHC21 and DHHC21

ZDHHC22 and DHHC22

ZDHHC23 and DHHC23

ZDHHC24 and DHHC24

The presence of ZDHHC gene transcripts in human blood cells and blood cell lines

Conclusions

Acknowledgements

Biochim. Biophys. Acta

Trends Cell Biol.

J. Biol. Chem.

Biochim. Biophys. Acta

Neuron

Methods

Biol. Psychiatry

J. Biol. Chem.

Trends Biochem. Sci.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Neuron

J. Lipid Res.

Cancer Genet. Cytogenet.

Brain Res. Mol. Brain Res.

J. Biol. Chem.

J. Biol. Chem.

Mol. Cell. Neurosci.

J. Lipid Res.

Curr. Biol.

Biochim. Biophys. Acta

Neurosci. Lett.

Am. J. Hum. Genet.

Cell. Signal.

Biochim. Biophys. Acta

Methods

Neurosci. Lett.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Bone

J. Biol. Chem.

Mol. Cell. Proteomics

Biochim. Biophys. Acta

Search for transmembrane protein in gastric cancer by the Escherichia coli ampicillin secretion trap: expression of DSC2 in gastric cancer with intestinal phenotype

J. Pathol.

Allelic loss of chromosomal arm 8p in breast cancer progression

Am. J. Pathol.

Paralemmin-1, a modulator of filopodia induction is required for spine maturation

Mol. Biol. Cell

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Mol. Membr. Biol.

Homozygous deletion and frequent allelic loss of chromosome 8p22 loci in human prostate cancer

Cancer Res.

Case–control study and transmission disequilibrium test provide consistent evidence for association between schizophrenia and genetic variation in the 22q11 gene ZDHHC8

Hum. Mol. Genet.

GABAA receptor associated proteins: a key factor regulating GABAA receptor function

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Human DHHC proteins: A spotlight on the hidden player of palmitoylation