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Jennifer Greaves, Kevin R Munro, Stuart C Davidson, Matthieu Riviere, Justyna Wojno, Terry K Smith, Nicholas C O Tomkinson, Luke H Chamberlain
S-acylation is a major posttranslational modification, catalyzed by the zinc finger DHHC domain containing (zDHHC) enzyme family. S-acylated proteins can be modified by different fatty acids; however, very little is known about how zDHHC enzymes contribute to acyl chain heterogeneity. Here, we used fatty acid-azide/alkyne labeling of mammalian cells, showing their transformation into acyl-CoAs and subsequent click chemistry-based detection, to demonstrate that zDHHC enzymes have marked differences in their fatty acid selectivity...
February 21, 2017: Proceedings of the National Academy of Sciences of the United States of America
Anindit Mukherjee, Zhijian Wang, Carol L Kinlough, Paul A Poland, Allison L Marciszyn, Nicolas Montalbetti, Marcelo D Carattino, Michael B Butterworth, Thomas R Kleyman, Rebecca P Hughey
The epithelial sodium channel (ENaC) has an important role in regulating extracellular fluid volume and blood pressure, as well as airway surface liquid volume and mucociliary clearance. ENaC is a trimer of three homologous subunits (α, β, and γ). We previously reported that cytoplasmic residues on the β (βCys-43 and βCys-557) and γ (γCys-33 and γCys-41) subunits are palmitoylated. Mutation of Cys that blocked ENaC palmitoylation also reduced channel open probability. Furthermore, γ subunit palmitoylation had a dominant role over β subunit palmitoylation in regulating ENaC...
March 10, 2017: Journal of Biological Chemistry
Keyong Du, Shoko Murakami, Yingmin Sun, Casey L Kilpatrick, Bernhard Luscher
Insulin-dependent translocation of glucose transporter 4 (Glut4) to the plasma membrane plays a key role in the dynamic regulation of glucose homeostasis. We recently showed that this process is critically dependent on palmitoylation of Glut4 at Cys-223. To gain further insights into the regulation of Glut4 palmitoylation, we set out to identify the palmitoyl acyltransferase (PAT) involved. Here we report that among 23 mammalian DHHC proteins, DHHC7 is the major Glut4 PAT, based on evidence that ectopic expression of DHHC7 increased Glut4 palmitoylation, whereas DHHC7 knockdown in 3T3-L1 adipocytes and DHHC7 KO in adipose tissue and muscle decreased Glut4 palmitoylation...
February 17, 2017: Journal of Biological Chemistry
Hitoshi Matakatsu, Seth S Blair, Richard G Fehon
The large protocadherin Fat functions to promote Hippo pathway activity in restricting tissue growth. Loss of Fat leads to accumulation of the atypical myosin Dachs at the apical junctional region, which in turn promotes growth by inhibiting Warts. We previously identified Approximated (App), a DHHC domain palmitoyltransferase, as a negative regulator of Fat signaling in growth control. We show here that App promotes growth by palmitoylating the intracellular domain of Fat, and that palmitoylation negatively regulates Fat function...
January 2, 2017: Journal of Cell Biology
Bo Zhou, Jian Zhong Lin, Dan Peng, Yuan Zhu Yang, Ming Guo, Dong Ying Tang, Xiaofeng Tan, Xuan Ming Liu
In many plants, architecture and grain yield are affected by both the environment and genetics. In rice, the tiller is a vital factor impacting plant architecture and regulated by many genes. In this study, we cloned a novel DHHC-type zinc finger protein gene Os02g0819100 and its alternative splice variant OsDHHC1 from the cDNA of rice (Oryza sativa L.), which regulate plant architecture by altering the tiller in rice. The tillers increased by about 40% when this type of DHHC-type zinc finger protein gene was over-expressed in Zhong Hua 11 (ZH11) rice plants...
January 2017: Plant Science: An International Journal of Experimental Plant Biology
Casey L Kilpatrick, Shoko Murakami, Mengyang Feng, Xia Wu, Rachnanjali Lal, Gong Chen, Keyong Du, Bernhard Luscher
The γ2 subunit of GABA type A receptors (GABAARs) is thought to be subject to palmitoylation by both Golgi-associated DHHC-type zinc finger protein (GODZ; also known as DHHC3) and its paralog Sertoli cell gene with a zinc finger domain-β (SERZ-β; DHHC7) based on overexpression of enzymes and substrates in heterologous cells. Here we have further investigated the substrate specificity of these enzymes by characterization of GODZ and SERZ-β knock-out (KO) mice as well as double KO (DKO) neurons. Palmitoylation of γ2 and a second substrate, growth-associated protein of 43 kDa, that is independently implicated in trafficking of GABAARs was significantly reduced in brain of GODZ KO versus wild-type (WT) mice but unaltered in SERZ-β KO mice...
December 30, 2016: Journal of Biological Chemistry
Baohui Zheng, Gopala K Jarugumilli, Baoen Chen, Xu Wu
Palmitoleoylation is a unique fatty acylation of proteins in which a monounsaturated fatty acid, palmitoleic acid (C16:1), is covalently attached to a protein. Wnt proteins are known to be palmitoleoylated by cis-Δ9 palmitoleate at conserved serine residues. O-palmitoleoylation plays a critical role in regulating Wnt secretion, binding to the receptors, and in the dynamics of Wnt signaling. Therefore, protein palmitoleoylation is important in tissue homeostasis and tumorigenesis. Chemical probes based on saturated fatty acids, such as ω-alkynyl palmitic acid (Alk-14 or Alk-C16 ), have been used to study Wnt palmitoleoylation...
November 3, 2016: Chembiochem: a European Journal of Chemical Biology
Jun Han, Hai Zhang, Sheng Wang, Jun Zhou, Yi Luo, Li-Hong Long, Zhuang-Li Hu, Fang Wang, Jian-Guo Chen, Peng-Fei Wu
Sulfhydryl compounds such as dithiothreitol (DTT) and β-mercaptoethanol (β-ME) are widely used as redox agents. Previous studies in our group and other laboratory have reported the effect of sulfhydryl compounds on the function of glutamate receptor, including plasticity. Most of these findings have focused on the N-methyl-D-aspartic acid receptor, in contrast, very little is known about the effect of sulfhydryl compounds on α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor (AMPAR). Here, we observed that DTT (100 μM), β-ME (200 μM) and L-cysteine (200 μM) significantly elevated the surface expression of AMPARs via reducing their palmitoylation in rat hippocampal slices in vitro...
November 2016: Neurochemical Research
Norihiko Yokoi, Yuko Fukata, Atsushi Sekiya, Tatsuro Murakami, Kenta Kobayashi, Masaki Fukata
UNLABELLED: Postsynaptic density (PSD)-95, the most abundant postsynaptic scaffolding protein, plays a pivotal role in synapse development and function. Continuous palmitoylation cycles on PSD-95 are essential for its synaptic clustering and regulation of AMPA receptor function. However, molecular mechanisms for palmitate cycling on PSD-95 remain incompletely understood, as PSD-95 depalmitoylating enzymes remain unknown. Here, we isolated 38 mouse or rat serine hydrolases and found that a subset specifically depalmitoylated PSD-95 in heterologous cells...
June 15, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Patricia Marie-Jeanne Lievens, Tatiana Kuznetsova, Gaga Kochlamazashvili, Fabrizia Cesca, Natalya Gorinski, Dalia Abdel Galil, Volodimir Cherkas, Natalia Ronkina, Juri Lafera, Matthias Gaestel, Evgeni Ponimaskin, Alexander Dityatev
The neural cell adhesion molecule (NCAM) mediates cell-cell and cell-matrix adhesion. It is broadly expressed in the nervous system and regulates neurite outgrowth, synaptogenesis, and synaptic plasticity. Previous in vitro studies revealed that palmitoylation of NCAM is required for fibroblast growth factor 2 (FGF2)-stimulated neurite outgrowth and identified the zinc finger DHHC (Asp-His-His-Cys)-containing proteins ZDHHC3 and ZDHHC7 as specific NCAM-palmitoylating enzymes. Here, we verified that FGF2 controlled NCAM palmitoylation in vivo and investigated molecular mechanisms regulating NCAM palmitoylation by ZDHHC3...
September 1, 2016: Molecular and Cellular Biology
Marilyn D Resh
Long, short and medium chain fatty acids are covalently attached to hundreds of proteins. Each fatty acid confers distinct biochemical properties, enabling fatty acylation to regulate intracellular trafficking, subcellular localization, protein-protein and protein-lipid interactions. Myristate and palmitate represent the most common fatty acid modifying groups. New insights into how fatty acylation reactions are catalyzed, and how fatty acylation regulates protein structure and function continue to emerge. Myristate is typically linked to an N-terminal glycine, but recent studies reveal that lysines can also be myristoylated...
July 2016: Progress in Lipid Research
Chwen L Tay, Matthew L Jones, Nicola Hodson, Michel Theron, Jyoti S Choudhary, Julian C Rayner
Palmitoylation is the post-translational reversible addition of the acyl moiety, palmitate, to cysteine residues of proteins, and is involved in regulating protein trafficking, localisation, stability and function. The DHHC protein family, named for their highly conserved Aspartate-Histidine-Histidine-Cysteine (DHHC) signature motif, is thought to be responsible for catalysing protein palmitoylation. Palmitoylation is widespread in all eukaryotes, including the malaria parasite, Plasmodium falciparum, where over 400 palmitoylated proteins are present in the asexual intraerythrocytic schizont stage parasites, including proteins involved in key aspects of parasite maturation and development...
April 6, 2016: Cellular Microbiology
Yuanwei Zhang, Qingqing Zheng, Congcong Sun, Jinxing Song, Lina Gao, Shizhu Zhang, Alberto Muñoz, Nick D Read, Ling Lu
Finely tuned changes in cytosolic free calcium ([Ca2+]c) mediate numerous intracellular functions resulting in the activation or inactivation of a series of target proteins. Palmitoylation is a reversible post-translational modification involved in membrane protein trafficking between membranes and in their functional modulation. However, studies on the relationship between palmitoylation and calcium signaling have been limited. Here, we demonstrate that the yeast palmitoyl transferase ScAkr1p homolog, AkrA in Aspergillus nidulans, regulates [Ca2+]c homeostasis...
April 2016: PLoS Genetics
Yuko Fukata, Tatsuro Murakami, Norihiko Yokoi, Masaki Fukata
Palmitoylation is an evolutionally conserved lipid modification of proteins. Dynamic and reversible palmitoylation controls a wide range of molecular and cellular properties of proteins including the protein trafficking, protein function, protein stability, and specialized membrane domain organization. However, technical difficulties in (1) detection of palmitoylated substrate proteins and (2) purification and enzymology of palmitoylating enzymes have prevented the progress in palmitoylation research, compared with that in phosphorylation research...
2016: Current Topics in Membranes
Wei Shi, Xueran Chen, Fen Wang, Ming Gao, Yang Yang, Zhaoxia Du, Chen Wang, Yao Yao, Kun He, Aijun Hao
In vertebrates, neural stem/progenitor cells (NSPCs) maintenance is critical for nervous system development and homeostasis. However, the molecular mechanisms underlying the maintenance of NSPCs have not been fully elucidated. Here, we demonstrated that zebrafish ZDHHC16, a DHHC encoding protein, which was related to protein palmitoylation after translation, was expressed in the developing forebrain, and especially in the telencephalon. Loss- and gain-of-function studies showed that ZDHHC16 played a crucial role in the regualtion of NSPCs proliferation during zebrafish telencephalic development, via a mechanism dependent on its palmitoyltransferase activity...
September 2016: Developmental Neurobiology
Tielin Zhou, Jing Li, Peiquan Zhao, Huijuan Liu, Deyong Jia, Hao Jia, Lin He, Yong Cang, Sharon Boast, Yi-Han Chen, Hélène Thibault, Marielle Scherrer-Crosbie, Stephen P Goff, Baojie Li
Protein palmitoylation regulates many aspects of cell function and is carried out by acyl transferases that contain zf-DHHC motifs. The in vivo physiological function of protein palmitoylation is largely unknown. Here we generated mice deficient in the acyl transferase Aph2 (Ablphilin 2 or zf-DHHC16) and demonstrated an essential role for Aph2 in embryonic/postnatal survival, eye development, and heart development. Aph2(-/-) embryos and pups showed cardiomyopathy and cardiac defects including bradycardia. We identified phospholamban, a protein often associated with human cardiomyopathy, as an interacting partner and a substrate of Aph2...
December 22, 2015: Proceedings of the National Academy of Sciences of the United States of America
Yaxiao Li, Rod Scott, James Doughty, Murray Grant, Baoxiu Qi
The Asp-His-His-Cys-Cys-rich domain-containing Protein S-Acyl Transferases (PATs) are multipass transmembrane proteins that catalyze S-acylation (commonly known as S-palmitoylation), the reversible posttranslational lipid modification of proteins. Palmitoylation enhances the hydrophobicity of proteins, contributes to their membrane association, and plays roles in protein trafficking and signaling. In Arabidopsis (Arabidopsis thaliana), there are at least 24 PATs; previous studies on two PATs established important roles in growth, development, and stress responses...
January 2016: Plant Physiology
Colin D Gottlieb, Sheng Zhang, Maurine E Linder
DHHC palmitoyltransferases catalyze the addition of the fatty acid palmitate to proteins on the cytoplasmic leaflet of cell membranes. There are 23 members of the highly diverse mammalian DHHC protein family, all of which contain a conserved catalytic domain called the cysteine-rich domain (CRD). DHHC proteins transfer palmitate via a two-step catalytic mechanism in which the enzyme first modifies itself with palmitate in a process termed autoacylation. The enzyme then transfers palmitate from itself onto substrate proteins...
December 4, 2015: Journal of Biological Chemistry
Małgorzata Prajer, Sebastian Tarcz
The thermosensitive ts111 mutant of Parameciun tetraurelia carries a recessive mutation which causes cell death after 2-8 divisions at the restrictive temperature of 35 degrees C. Expression at 35 degrees C induces disassembly of the infraciliary lattice (ICL). In this study, we found that the ts111 mutation also results in significant abnormalities in the number and structure of contractile vacuole complexes (CVCs) and in their functioning at the restrictive temperature. In order to characterize the ts111 gene, the complementation cloning was performed by microinjection into the macronucleus of an indexed genomic DNA library...
2015: Folia Biologica (Krakow)
Michael Veit, Stefanie Siche
Covalent attachment of saturated fatty acids (palmitate and stearate) to hemagglutinin (HA) of influenza virus is a protein modification essential for viral replication. The enzymes catalysing acylation of viral proteins have not been identified, but likely candidates that acylate cellular substrates are members of a protein family that contain a DHHC (Asp-His-His-Cys) cysteine-rich domain. Since 23 DHHC-proteins with distinct, only partly overlapping substrate specificities are present in humans, only a few of them might acylate HA in airway cells of the lung...
December 8, 2015: Vaccine
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