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CILP-2 is a novel secreted protein and associated with insulin resistance

Journal of Molecular Cell Biology 2019年第12期11卷 1083-1094页

作者： Tong Wu Qin Zhang Shaobo Wu Wenjing Hu Tingting Zhou Ke Li Dongfang Liu Harvest FGu Hongting Zheng Zhiming Zhu Ling Li and Gangyi Yang Department of Endocrinology The Second Affiliated HospitalChongqing Medical UniversityChongqing Clinical Research Center for GeriatricsChongqingChina Key Laboratory of Diagnostic Medicine(Ministry of Education)and Department of Clinical Biochemistry College of Laboratory MedicineChongqing Medical UniversityChongqingChina Center for Pathophysiology School of Basic Medicine and Clinical PharmacyChina Pharmaceutical UniversityNanjingChina Department of Endocrinology Xinqiao HospitalThird Military Medical UniversityChongqingChina Department of Hypertension and Endocrinology Daping HospitalThird Military Medical UniversityChongqing Institute of HypertensionChongqingChina

Genetic association studies have implicated that cartilage intermediate layer protein 2 (CILP-2) confers the risk susceptibility for type 2 diabetes (T2DM). However, it is still unknown whether CILP-2 is involved in the regulation of glucose homeostasis and insulin resistance (IR). In the current study, we initially observed that CILP-2 as a secreted protein was detected in both conditioned medium and lysates of cells transfected with an overexpressed vector. We then found that circulating CILP-2 levels had a progressive increase from normal to impaired glucose tolerance (a pre-diabetic status) and then to diabetes, which was correlated positively with waist-to-hip ratio, triglyceride, fasting blood glucose, 2-h blood glucose after glucose overload, HbA1c, fasting insulin, 2-h plasma insulin after glucose overload, and homeostasis model assessment of insulin resistance but negatively with HDL-C. CILP-2 expression was increased in the liver and muscle but decreased in adipose tissues of obese mice or T2DM patients. Furthermore, we demonstrated that CILP-2 circulating levels were affected by OGTT and Exenatide. CILP-2 overexpression resulted in impaired glucose tolerance and hepatic IR in vivo and increased PEPCK expression whereas suppressed phosphorylation of insulin receptor and Akt kinase in vitro. Based on these findings, we have identified a direct interaction between CILP-2 and PEPCK and suggested that CILP-2 plays an important role in the regulation of hepatic glucose production.

关键词： CILP-2 secreted protein insulin resistance type 2 diabetes

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Functional research on a secreted protein SsCFEM1 in Sclerotinia sclerotiorum

Functional research on a secreted protein SsCFEM1 in Sclerot...

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中国植物病理学会2019年学术年会

作者： Ji Xu Liu Jinliang Zhang Yanhua Zhang Xianghui Pan Hongyu College of Plant Sciences Jilin University

Sclerotinia sclerotiorum（Lib.） de Bary is a broad host range plant pathogen and causes lots of loss in the worldwide scale every year.Sclerotinia sclerotiorum can form the infection structure,named comprounded appressorium,in order to adhere and penetrate the host plant during pathogenic processes.In addition,Sclerotinia sclerotiorum is able to synthesize oxalic acid and cell wall degrading enzymes,which play important role in resisting host defense reaction and accelerating pathogenicity.CFEM（common in several fungal extracellular membrane proteins） domain is unique in fungi and contains eight conserved cysteine residues.InMagnaporthe oryzae,the CFEM domain of seven-transmembrane protein Pth11,a significant G-protein-coupled receptor,is necessary for proper development of appressoria,appressorialike structures and pathogenicity.However,the underlying mechanisms by which CFEM domain-containing proteins act remain largely unknown,especially in broad-hostrange necrotrophic pathogen 5.Sclerotiorum.The transcript level of SsCFEM1 increased highly at 24 hours post inoculation（hpi） and then maintained at a high level expression during late stages（24～48 hpi）,indicating that SsCFEM1 may be involved in infection of S.sclerotiorum.In order to futher investigate the role of SsCFEM1 in S.Sclerotiorum,the knock-out mutant was obtained by PEG-mediated protoplast transformation.The knock-out mutant showed no significant changes on mycelial growth,colony morphology and sclerotial development compared with wild type strain.Although the knock-out mutant can form normal comprounded appressorium,the number of it is significantly reduced.Virulence of knock-out mutants was significant declined on detached leaves on Nicotiana benthamiana,suggesting that SsCFEM1 is involved in the virulence of S.sclerotiorum.Therefore,our data suggested that SsCFEM1 contributes to virulence and comprounded appressorium production.

关键词： Sclerotinia sclerotiorum secreted protein SsCFEM1

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Endothelial METRNL determines circulating METRNL level and maintains endothelial function against atherosclerosis

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Acta Pharmaceutica Sinica B 2023年第4期13卷 1568-1587页

作者： Sili Zheng Zhiyong Li Jie Song Pin Wang Jian Xu Wenjun Hu Yi Shi Qi Qi Zhuwei Miao Yunfeng Guan Chaoyu Miao Department of Pharmacology Second Military Medical University/Naval Medical UniversityShanghai 200433China Changhai Hospital Second Military Medical University/Naval Medical UniversityShanghai 200433China Zhongshan Hospital Fudan UniversityShanghai 200032China

METRNL is a recently identified secreted protein with emerging functions.This study is to find major cellular source of circulating METRNL and to determine METRNL novel function.Here,we show METRNL is abundant in human and mouse vascular endothelium and released by endothelial cells using endoplasmic reticulum-Golgi apparatus pathway.By creating endothelial cell-specific Metrnl knockout mice,combined with bone marrow transplantation to produce bone marrow-specific deletion of Metrnl,we demonstrate that most of circulating METRNL(approximately 75%)originates from the endothelial cells.Both endothelial and circulating METRNL decrease in atherosclerosis mice and patients.By generating endothelial cell-specific Metrnl knockout in apolipoprotein E-deficient mice,combined with bone marrow-specific deletion of Metrnl in apolipoprotein E-deficient mice,we further demonstrate that endothelial METRNL deficiency accelerates atherosclerosis.Mechanically,endothelial METRNL deficiency causes vascular endothelial dysfunction including vasodilation impairment via reducing eNOS phosphorylation at Ser1177 and inflammation activation via enhancing NFκB pathway,which promotes the susceptibility of atherosclerosis.Exogenous METRNL rescues METRNL deficiency induced endothelial dysfunction.These findings reveal that METRNL is a new endothelial substance not only determining the circulating METRNL level but also regulating endothelial function for vascular health and disease.METRNL is a therapeutic target against endothelial dysfunction and atherosclerosis.

关键词： METRNL secreted protein Endothelium Endothelial function Bone marrow Atherosclerosis

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Histone H3K27me3 methylation regulates the expression of secreted proteins distributed at fast-evolving regions through transcriptional repression of transposable elements

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Journal of Integrative Agriculture 2023年第10期22卷 3059-3068页

作者： XIE Jia-hui TANG Wei LU Guo-dong HONG Yong-he ZHONG Zhen-hui WANG Zong-hua ZHENG Hua-kun State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops College of Plant ProtectionFujian Agriculture and Forestry UniversityFuzhou 350002P.R.China Fujian Universities Key Laboratory for Plant Microbe Interaction College of Life SciencesFujian Agriculture and Forestry UniversityFuzhou 350002P.R.China Fuzhou Institute of Oceanography Minjiang UniversityFuzhou 350108P.R.China National Engineering Research Center of JUNCAO Technology College of Life ScienceFujian Agriculture and Forestry UniversityFuzhou 350002P.R.China

The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation of these genes remain largely unknown.Here,through comparative transcriptome and chromatin immunoprecipitation sequencing(ChIP-seq)analyses of the methyltransferase PoKmt6 in rice blast fungus Pyricularia oryzae(syn.Magnaporthe oryzae),we found that PoKmt6-mediated H3K27me3 deposition was enriched mainly at fast-evolving regions and contributed to the silencing of a subset of secreted proteins(SP)and transposable element(TE)families during the vegetative growth of P.oryzae.Intriguingly,we observed that a group of SP genes,which were depleted of H3K27me3 modification,could also be silenced via the H3K27me3-mediated repression of the nearby TEs.In conclusion,our results indicate that H3K27me3 modification mediated by PoKmt6 regulates the expression of some SP genes in fast-evolving regions through the suppression of nearby TEs.

关键词： secreted protein transposable elements fast-evolving regions H3K27me3

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Plant Diseases and Pests 2017年第2期8卷 26-33页

作者： Wang Hui Su Yuan Liu Chao Wang Yi Yang Jing Li Chengyun State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan Yunnan Agricultural University

The structure similarity of secreted proteins in rice blast fungus Magnaporthe oryzae and its host Oryza sativa was analyzed. One thousand two hundred and forty one proteins were predicted as secreted proteins using four algorithms based on 11 074 proteins in genome of M. oryzae. One hundred and forty six secreted proteins（ 11. 8% of M. oryzae secretome） were aligned with 116 rice proteins（ 0. 21% of 56 278 rice proteins） using BLAST search on rice genome. One hundred sixteen rice similar proteins participated in rice cell wall modification（ cell wall associated enzymes） and signal transduction（ proteases）. These results imply that both cell wall involved proteins and signal transduction are probably hijacks pathway between host pants and pathogenic fungi. Because these proteins are highly conserved among fungi and plants,the express patterns of these protein coding genes during the interaction process are valuable to study in detail.

关键词： Blast fungus Rice secreted protein Structure similarity

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Genes encoding a group of related small secreted proteins from the gut of Hessian fly larvae [Mayetiola destructor （Say）]

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Insect Science 2006年第5期13卷 339-348页

作者： MING-SHUN CHEN XIANG LIU YU-CHENG ZHU JOHN C. REESE GERALD E. W ILDE USDA-ARS Plant Science and Entomology Research UnitUSA Department of Entomology Kansas State University Manhattan KansasUSA USDA-ARS-JWDSRC Stoneville Minnesota USA

A group of related genes has been isolated and characterized from the gut of Hessian fly larvae [Mayetiola destructor （Say）]. Members in this group appear to encode proteins with secretary signal peptides at the N-terminals. The mature putative proteins are small, acidic proteins with calculated molecular masses of 14.5 to 15.3 kDa, and isoelectric points from 4.56 to 4.88. Northern blot analysis revealed that these genes are expressed predominantly in the gut of Hessian fly larvae and pupae. Two related genes, GIOK1 and GIOK2, were isolated as tandem repeats. Both genes contain three exons and two introns. The intron/exon boundaries were conserved in terms of amino acid encoding, suggesting that they arose by gene duplication. The fact that the frequency of this group of clones in a gut cDNA library higher than that of total cDNA clones encoding digestive enzymes suggested that this group of proteins may perform an important function in the gut physiology of this insect. However, the exact functions of these proteins are as yet known since no sequence similarity could be identified between these proteins and any known sequences in public databases using standard methods.

关键词： Hessian fly gut Mayetiola destructor secreted protein wheat

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Candidate effector proteins of the necrotrophic apple canker pathogen Valsa mali can suppress BAX-induced PCD

Candidate effector proteins of the necrotrophic apple canker...

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中国植物病理学会第十二届青年学术研讨会

作者： Zhengpeng Li Zhiyuan Yin Yanyun Fan Ming Xu Zhensheng Kang Lili Huang State Key Laboratory of Crop Stress Biology for Arid Areas Northwest A&F University China-Australia Joint Research Centre for Abiotic and Biotic Stress Management Northwest A&F University

Canker caused by the Ascomycete Valsa mali is the most destructive disease of apple in Eastern Asia,resulting in yield losses of up to 100%.This necrotrophic fungus induces severe necrosis on apple,eventually leading to the death of the whole tree.Identification of necrosis inducing factors may help to unravel the molecular bases for colonization of apple trees by V.mali.As a first step toward this goal,we identified and characterized the V.mali repertoire of candidate effector proteins（CEPs）.In total,193 secreted proteins with no known function were predicted from genomic data,of which 101 were V.mali-speciiic.Compared to non-CEPs predicted for the V.mali secretome.CEPs have shorter sequence length and a higher content of cysteine residues.Based on transient over-expression in Nicotiana benthamiana performed for 70 randomly selected CEPs,seven V.mali Effector proteins（VmEPs）were shown to significantly suppress BAX-induced PCD.Furthermore,targeted deletion of VmEP1 resulted in a significant reduction of virulence.These results suggest that V.mali expresses secreted proteins that can suppress PCD usually associated with effector-triggered immunity（ETI）.ETI in turn may play an important role in the V.mali-apple interaction.The ability of V.mali to suppress plant ETI sheds a new light onto the interaction of a necrotrophic fungus with its host plant.

关键词： apple Valsa canker secreted protein cell death virulence factor plantfungus interaction

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Intestine metrnl acts as a local regulator released into gut lumen and maintaining gut antimicrobial peptides

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中国药理学与毒理学杂志 2016年第10期30卷 1092-1092页

作者： LI Zhi-yong FAN Mao-bing QU Yi ZHENG Si-li SONG Jie MIAO Chao-yu Department of Pharmacology Second Military Medical University

OBJECTIVE Metrnl is a novel secreted protein with limited researches.In this study,we investigated metrnl tissue expression pattern in humans,and exploredthe possible role of its highest expression using animal models.METHODS We examined metrnl tissue expression pattern in a human tissue microarray containing 19types of tissues from 69 donors,and verified the highest expression in fresh human and mouse tissues.We then created an animal model of cell-specific knockout mice to study the role of metrnl.RESULTS Metrnl was the highest expressed in human gastrointestinal tract,and specifical y expressed in the intestinal epithelium.Consistently,Metrnl expression was also the highest expressed in mouse gastrointestinal tract among the detected tissues of 14 types.We developed intestinal epithelial cellspecific metrnl knockout mice with Vil in-Cre.In this animal model,metrnl levels displayed a statistically significant reduction in gut fluid,but not in blood serum.This cell specific deletion of metrnl did not affect body weight,food intake,blood glucose,colon length and histology,intestinal permeability,mucus production and mucin 2 expression under physiological conditions,but markedly reduced the expression of antimicrobial peptides,such as regenerating islet-derived 3 gamma and lactotransferrin.CONCLUSION Metrnl is rich in intestinal epithelial cells of humans and mice,mainly contributing to local gut metrnl level,and less affecting systemic circulating metrnl level.Metrnl plays a role in maintaining gut antimicrobial peptides.

关键词： metrnl secreted protein human expression pattern intestine epithelial cel antimicrobial peptide

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Computational Analysis of Signal Peptide-Dependent secreted proteins in Saccharomyces cerevisiae

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Agricultural Sciences in China 2006年第3期5卷 221-227页

作者： YANG Jing LI Cheng-yun WANG Yun-yue ZHU You-yong LI Jin-bin HE Xia-hong ZHOU Xiao-gang LIU Lin YE Yan-fen Key Laboratory for Agricultural Biodiversity for Pest Management Ministry of Education/~unnan Agricultural University Kunming 650223 P.R.China Plant Protection Research Institute Yunnan Academy of Agricultural Sciences Kunming 650201 P.R.China Biotechnology Research Institute Yunnan Academy of Agricultural Sciences Kunming 650223 P.R.China

Computer based software such as the SignalP v3.0, TargetP v1.01, big-PI predictor and TMHMM v2.0 were combined to predict the signal peptides, and the signal peptide-dependent secreted proteins among the 6 700 ORFs in genome of Saccharomyces cerevisiae. The results showed that 163 proteins were the secreted ones containing signal peptides, and they were secreted via Sec pathway. Among the 163 predicted secreted proteins, the signal peptides of 47 secreted proteins included only the H-domain and C-domain, without N-domain, but the signal peptides of other 116 secreted proteins included all the three domains. There were differences in the constitution of signal peptides between the secreted proteins of S. cerevisiae and of Candida albicans, but the length and amino acids types of their signal peptides were similar in general. Few of the same signal peptides occurred in the secreted proteins of S. cerevisiae genome, and the homology could be compared among the secreted proteins with the same signal peptides. The BLAST 2 SEQUENECES and CLUSTAL W were used to align the two protein sequences and multi-protein sequences, respectively. The alignment result indicated that homology of these sequences with the same signal peptide was very highly conservative in amino acid of complete gene. The effect of the signal peptides in S. cerevisia on expression of foreign eukaryotic secreted proteins is discussed in this paper.

关键词： Saccharomyces cerevisia secreted protein signal peptide secretory pathway

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Synaptic vesicle proteins are transported to axon terminals by different vesicle precursors

Synaptic vesicle proteins are transported to axon terminals ...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Chang-Lin LI,Xu ZHANG~* Institute of Neuroscience and State Key Laboratory of Neuroscience,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,Shanghai 200031,China

Objective The central afferent terminals of the primary sensory neurons release excitatory neurotransmitter glutamate and neuropeptides such as substance P and calcitonin gene-related peptide(CGRP) via exocytosis of synaptic vesicles and large dense-core vesicles(LDCVs),respectively.In response to stimulation,the intracellular Ca is increased, which triggers exocytosis of the synaptic vesicles and LDCVs.Some neuropeptides are synthesized in endoplasmic reticulum,processed in the Golgi complex,and sorted to LDCVs in cell body of small dorsal root ganglion(DRG) neurons. After budding off from the trans-Golgi network(TGN),LDCVs are delivered to axonal terminals for secretion.Although synaptic vesicles are suggested by some studies to be formed in TGN,they are proposed to be assembled and recycled locally in presynaptic terminals.And synaptic vesicle proteins are in the need to be delivered from soma to axon terminals via synaptic vesicle precursors.However,the transportation of these synaptic vesicle precursors to axon terminals is not clear.And whether there are the contents inside synaptic vesicle precursors remains unknown.Methods The localization of synaptic vesicle proteins(VGluT2,VAMP2,Synapsin and Synaptoporin) was identified by immunolabeling and vesicle immunoisolation.Subcellular fractionation with a sucrose gradient was used to separate different vesicle populations in rat spinal dorsal horn.And the secreted protein was determined by proteomic analysis of VGluT-2 containing fraction.The concentration of glutamate in VGluT2-containing synaptic vesicle precursors was determined by HPLC.Results(1) VGluT2,VAMP2 and synaptoporin were mainly present in small translucent vesicles from dorsal roots,but not in LDCVs indicated by CGRP.(2) VAMP2 was partly localized in VGluT2-positive vesicle precursors in dorsal roots,which was confirmed by the results of vesicle immunoisolation.(3) Synapsin was only partly localized in VGluT2-containing synaptic vesicle precursors in dorsal roots.(4) Proteomic analysis showed that

关键词： synaptic vesicle synaptic vesicle precursor LDCV secreted protein glutamate

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