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Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis

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Zoological Research 2019年第3期40卷 198-204页

作者： Fen Zhang Zhi-Lai Guo Yan chen Li Li Hai-Ning Yu Yi-Peng Wang Department of Pharmaceutical Sciences College of Pharmaceutical SciencesSoochow UniversitySuzhou Jiangsu 215123China Department of Bioscience and Biotechnology Dalian University of TechnologyDalian Liaoning 116023China School of Life Sciences Guizhou Normal UniversityGuiyang Guizhou 550001China

Rana kunyuensis is a species of brown frog that lives exclusively on Kunyu Mountain,Yantai,china.In the current study,a 279-bp cDNA sequence encoding a novel antimicrobial peptide (AMP),designated as amurin-9KY,was cloned from synthesized double-strand skin cDNA of R.kunyuensis.The amurin-9KY precursor was composed of 62 amino acid (aa) residues,whereas the mature peptide was composed of 14 aa and contained two cysteines forming a C-terminal heptapeptide ring (Rana box domain) and an amidated c-terminus.These structural characters represent a novel amphibian AMP family.Although amurin-9KY exhibited high similarity to the already identified amurin-9AM from R.amurensis,little is known about the structures and activities of amurin-9 family AMPs so far.Therefore,amurin-9KY and its three derivatives (amurin-9KY1-3) were designed and synthesized.The structures and activities were examined to evaluate the influence of C-terminal amidation and the heptapeptide ring on the activities and structure of amurin-9KY..Results indicated that C-terminal amidation was essential for antimicrobial activity,whereas both C-terminal amidation and the heptapeptide ring played roles in the low hemolytic activity.circular dichroism (cD) spectra showed that the four peptides adopted an α-helical conformation in THF/H2O (v/v 1∶1) solution,but a random coil in aqueous solution.Elimination of the C-terminal heptapeptide ring generated two free cysteine residues with unpaired thiol groups,which greatly increased the concentration-dependent anti-oxidant activity.Scanning electron microscopy (SEM) was also performed to determine the possible bactericidal mechanisms.

关键词： Antimicrobial peptides Rana kunyuensis',Amurin-9KY Heptapeptide ring C-terminal amidation Structure activity relati on ship

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Modulation of voltage-gated potassium Kv2.1 via the cytoplasmic c terminal

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Neural Regeneration Research 2011年第7期6卷 553-557页

作者： Man Jin Peiyuan Lu Department of Neurology Hebei General Hospital Shijiazhuang 050051 Hebei Province China

Voltage-gated potassium channels comprise 12 subtypes （Kv1-Kv12）. Kv2.1, which is expressed in most mammalian central neurons, provides the majority of delayed-rectifier K＋ current in cortical and hippocampal pyramidal neurons, and plays an especially prominent role in repolarizing membrane potential, as well as in facilitation of exocytosis. Kv2.1-encoded K＋ efflux is essential for neuronal apoptosis programming. The human form of the Kv2.1 potassium channel contains large intracellular regions. The cytoplasmic C-terminal plays a key role in modulating Kv2.1 gating. The present manuscript summarized Kv2.1 structure and modulation in neurons and analyzed the roles of the cytoplasmic C-terminal.

关键词： Kv2.1 C-terminal N-terminal N/c interaction voltage-dependent gating

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MALDI-TOF-TOF-MS对完整蛋白质c端序列的直接测定

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分析测试学报 2008年第S1期27卷 13-14页

作者：刘炳玉薛燕刘锋李萍王红霞王鸿丽魏开华国家生物医学分析中心北京100850

C-terminal sequencing is important for characterized the primary structure of protein and polypeptides and it must do for quality control of recombination protein drugs.A method of MALDI-TOF-TOF-MS with in source decay(ISD) to measure C-terminal sequence of protein was established.This method was applied successfully to measure C-terminal sequence of Neuregulin,TP-Ⅱ,ubiquitin,and myoglobin,respectively and 10,24,24 and 36 amino acid residue of C-terminal were measured.

关键词： C-terminal sequencing MALDI-TOF-TOF-MS ISD

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Magnesium regulates the Kcnql/Kcnel channel function via the C-terminal of Kcnel subunit

Magnesium regulates the Kcnql/Kcnel channel function via the...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： Zhi-Jun SHEN Ion channel Disease Lab,Department of Physiology and Pharmacology,Key Lab of Development and Disease Related Genes,School of Basic Medical Science,Southeast University School of Medicine,Nanjing 210009,china

＜正＞The voltage-dependent potassium channel Kv7.1 is a tetramer composed of Kcnq1 a and Kcnel b subunits.It has been found functionally expressed in heart,inner ear,kidney and colons and mediating depolarization in the cardiac myocytes and potassium secretion into endolymph in the inner ear epithelial cells and assisting chloride homeostasis in epithelium of kidney and colons under physiological condition.Regulatory mechanisms of the channel function via adrenergic and purinergic receptors are well documented.Inherited mutations found in both kcnql and kcnel gene have been directly linked to clinical phenotypes of dominant form of Long QT syndrome with hearing loss（Jervell-Lange-Neilson Syndrome） and recessive form of Long QT syndrome without hearing loss（Romano-Word Syndrome）,which could be duplicated in experimental animals with gene engineering of knock-out and/or transgenic techniques.Here,we report that the intracellular magnesium is critical to the physiological function of the Kcnq 1/Kcne 1 channel.Using native inner ear epithelial cells,we recorded a voltage-dependent slowly activating current from the apical membrane via a macropatch on-cell configuration.The current was potassium selective and sensitive to the specific blocker of the Kcnq1/Kcne1 channel,chromano1 293B.Under excised or conventional whole cell configurations,we found that intracellular magnesium at assumed physiological levels was inhibitory to the function of the channel.The current was gradually running down upon excising or establishing the whole cell,and totally disappeared within 2-3 min.Adding ATP or Phosphatase inhibitor did not prevent the run-down.Elimination of magnesium in the background of physiological calcium levels could not only well keep the channel alive,but also significantly boosted the current 5-10 folds of its previous levels.Furthermore,the magnesium-free condition changed the voltage-dependency of the channel.It appeared that the channel carried two components under the magnesium-free condition;an instantaneous and

关键词： Kcnql/Kcnel channel intracellular magnesium C-terminal

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Molecular Mechanism of the Specificity of Protein Import into chloroplasts and Mitochondria in Plant cells

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Molecular Plant 2019年第7期12卷 951-966页

作者： Dong Wook Lee Sumin Lee Junho Lee Seungjin Woo Md.Abdur Razzak Alessandro Vitale Inhwan Hwang Division of Integrative Biosciences and Biotechnology Pohang University of Science and Technology Pohang 37673 Korea Department of Life Sciences Pohang University of Science and Technology Pohang 37673 Korea Istituto di Biologia e Biotecnologia Agraria Consiglio Nazionale Delle Ricerche Milano Italy

Plants possess both types of endosymbiotic organelles, chloroplasts and mitochondria. Transit peptides and presequences function as signal sequences for specific import into chloroplasts and mitochondria, respectively. However, how these highly similar signal sequences confer the protein import specificity remains elusive. Here, we show that mitochondrial- or chloroplast-specific import involves two distinct steps, specificity determination and translocation across envelopes, which are mediated by the N-terminal regions and functionally interchangeable C-terminal regions, respectively, of transit peptides and presequences. A domain harboring multiple-arginine and hydrophobic sequence motifs in the N-terminal regions of presequences was identified as the mitochondrial specificity factor. The presence of this domain and the absence of arginine residues in the N-terminal regions of otherwise common targeting signals confers specificity of protein import into mitochondria and chloroplasts, respectively. AtToc159, a chloroplast import receptor, also contributes to determining chloroplast import specificity. We propose that common ancestral sequences were functionalized into mitochondrial- and chloroplast-specific signal sequences by the presence and absence, respectively, of multiple-arginine and hydrophobic sequence motifs in the N-terminal region.

关键词： transit peptide presequence protein import into chloroplasts and mitochondria N-terminal specificity domain C-terminal common translocation domain import specificity determination

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The real-time second-derivative spectral analysis of c-terminus of engineering product by RP-HPLc

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chinese Science Bulletin 1995年第23期40卷 1997-2001页

作者：张丽华屠红旻邵晓霞吴高德徐来根夏其昌 Shanghai Institute of Biochemistry Chinese Academy of Sciences Shanghai 200031 China

N-terminal and C-terminal analysis of genetic engineering recombinant proteins are two criteria of determining their primary structure. The N-terminal sequencing relies heavily upon the Edman degradation method, which has been automated with commercially available sequencers. The maximum number of residues which may be placed in sequence in a single run is about 50. In the present decade, improvements in this technique made it

关键词： second-derivative spectrum C-terminal analysis new-type IL-2 (Ser-125) hEGF.

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EVI1 Mediated Stimulation of 3T3-L1 Preadipocyte Differentiation Is ctBP Dependent

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Yangtze Medicine 2017年第3期1卷 133-147页

作者： M. J. Ireland M. Al-Hasan J. A. craft A. Graham c. Bartholomew Department of Life Sciences School of Health & Life Sciences Glasgow Caledonian University City Campus Glasgow Scotland

Myelodysplasia syndrome 1 (MDS1) and Ecotropic viral integration site 1 (EVI1) complex (MEcOM) locus encode multiple isoforms of the EVI1 protein that are essential for normal vertebrate development and when inappropriately expressed play a significant role in malignancy and in particular leukaemias. However, the function of individual EVI1 isoforms is not fully understood. Recently, EVI1 or PRDM3, which is structurally closely related to the brown adipose tissue determining factor PRDM16, was shown to be required for differentiation of adipocytes. In this study, we show that 3T3-L1 preadipocytes sustain expression of all Evi1 isoforms examined, including Mds1-Evi1, Evi1FL, Evi1Δ324, Evi1FL + 9 and Evi1Δ105 throughout the adipogenesis differentiation programme. We also show that differentiation markers are enhanced by enforced expression of either Evi1, Evi1FL + 9 or Evi1Δ105 isoforms. Interestingly 3T3-L1 differentiation markers are also moderately enhanced by enforced expression of Evi1Δ324, which lacks part of the N-ter-minal zinc finger domain (ZF1), demonstrating a biological activity for this particular isoform. Enforced expression of an Evi1 mutant lacking C-terminal binding protein (ctBP) co-repressor protein binding activity fails to stimulate 3T3-L1 differentiation markers and may have dominant negative activity, causing partial inhibition of this developmental programme. These studies show that multiple EVI1 isoforms are expressed in adipocytes and can stimulate adipogenic markers in a manner that is partially independent of the ZF1 DNA binding domain but fully dependent upon interaction with co-repressor ctBP proteins.

关键词： MEcOM PRDM3 EVI1 Isoforms C-terminal Binding Proteins Adipogenesis

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