Thermostability of Doubly Glycosylated Recombinant Lysozyme
スポンサーリンク
概要
- 論文の詳細を見る
We prepared a lysozyme mutant(Q4lS/R61S)introducing Asn-type glycosylation signal sites by yeast expression system. On purification by cation exchange column at pH 7, three fractions were obtained. Peptide mapping and mass-spectrometry showed the fractions were the derivatives glycosylated at both Asn39 and Asn59, at only Asn39, and not glycosylated. It was revealed that the processing of Asn-linked oligosaccharide at Asn39 and Asn59 occurred independently in yeast cells. The denaturation temperatures of these derivatives by differential scanning calorimetry were 76.0, 68.8, and 67.5 ℃ at pH 3, respectively. The stabilization of glycosylated lysozyme depends on the degree of glycosylation. We concluded that stabilized proteins can be constructed by glycosylation at proper sites. Thermodynamic stabilization by the artificial double glycosylations on a protein has not yet been reported.
- 社団法人日本薬学会の論文
- 2001-10-01
著者
-
Ueda T
Department Of Chemistry And Biotechnology Graduate School Of Engineering The University Of Tokyo
-
Ueda T
Graduate School Of Pharmaceutical Sciences Kyushu University
-
IMOTO Taiji
Faculty of Biotechnology and Life Science, Sojo University
-
UEDA Tadashi
Graduate School of Pharmaceutical Sciences, Kyushu University
-
IMOTO Taiji
Graduate School of Pharmaceutical Sciences, Kyushu University
-
HASHIMOTO Yoshio
Graduate School of Pharmaceutical Sciences, Kyushu University
-
MUNEMURA Osamu
Graduate School of Pharmaceutical Sciences, Kyushu University
-
MASUMOTO Kiyonari
Graduate School of Pharmaceutical Sciences, Kyushu University
-
Masumoto Kiyonari
Graduate School Of Pharmaceutical Sciences Kyushu University
-
Imoto T
Faculty Of Biotechnology And Life Science Sojo University
-
Imoto Taiji
Graduate School Of Pharmaceutical Sciences
-
Imoto Taiji
Graduate School Of Pharmaceutical Science Kyushu University
-
Ueda Takuya
Department Of Chemistry And Biotechnology Graduate School Of Engineering The University Of Tokyo
-
上田 卓也
東京大学大学院新領域創成科学研究科メディカルゲノム専攻
-
Ueda Tadashi
Graduate School Of Pharmaceutical Sciences Kyushu Univ.
-
Ueda Tadashi
Graduate School Of Pharmaceutical Sciences
-
Imoto Taiji
九州大学 薬学研究院
-
Hashimoto Y
Graduate School Of Pharmaceutical Sciences Kyushu University
-
Hashimoto Yuichi
Institute Of Molecular & Cellular Biosciences University Of Tokyo
-
Hashimoto Yuichi
Institute Of Molecular And Cellular Biosciences The Universityv Of Tokyo
-
Munemura Osamu
Graduate School Of Pharmaceutical Sciences Kyushu University
-
Hashimoto Y
Institute Of Molecular & Cellular Biosciences The University Of Tokyo
-
Hashimoto Yoshio
Graduate School Of Pharmaceutical Sciences Kyushu University
関連論文
- 哺乳類ミトコンドリアにおける新規翻訳因子の発見
- Coordination of Divalent Metal Cation to Amide Group to Form Adduct Ion in FAB Mass Spectrometry: Implication of Zn^ in Enzymatic Hydrolysis of Amide Bond
- 蛋白質解析装置 無細胞蛋白質合成系:大腸菌由来完全再構成系PURESYSTEM (バイオ高性能機器・新技術利用マニュアル) -- (各種解析装置の原理と使用例)
- 無細胞タンパク質合成系へのピュアアプローチ
- Technology 無細胞タンパク質合成系の新展開
- 翻訳に必須な因子の精製と再構成による生体外タンパク質合成系(Purified System)の構築
- CCA末端修復酵素の反応機構の解析
- Crystal Structures of K33 Mutant Hen Lysozymes with Enhanced Activities
- 3P-017 変性条件下における抗体λ6鎖の残存構造がアミロイド線維形成に及ぼす影響(蛋白質・物性(3),第46回日本生物物理学会年会)
- Analysis of Internal Motions of RNase T1 Complexed with a Productive Substrate Involving ^N NMR Relaxation Measurements
- Characterization of Refolded Hen Lysozyme Variant Lacking Two Outside Disulfide Bonds
- Fluctuations in Free or Substrate-Complexed Lysozyme and a Mutant of It Detected on X-Ray Crystallography and Comparison with Those Detected on NMR
- Thermostability of Doubly Glycosylated Recombinant Lysozyme
- Expression and Characterization of Human Rheumatoid Factor Single-Chain FV
- Construction of the Single-Chain Fv from 196-14 Antibody toward Ovarian Cancer-Associated Antigen CA125
- Detection of a Local Interaction of Hen Lysozyme under Highly Denaturing Conditions Using Chemically ^C-Enriched Methionine Resonance
- An Improved Method for Preparing Lysozyme with Chemically ^C-Enriched methionine Residues Using 2-Aminothiophenol as a Reagent of Thiolysis
- Analysis of the Stability of Mutant Lysozymes at Position 15 Using X-Ray Crystallography
- Influence of Mutations of the N-Cap Residue, Gly4, on Stability and Structure of Hen Lysozyme
- Analysis of the Stabilization of Hen Lysozyme by Helix Macrodipole and Charged Side Chain Interaction
- Situation of Monomethoxypolyethylene Glycol Covalently Attached to Lysozyme
- Analysis of the Transition State in the Unfolding of Hen Lysozyme by Introduction of Gly-Pro and Pro-Gly Sequences at the Same Site
- Stabilization of Lysozyme by Introducing N-Glycosylation Signal Sequence
- A Mutation Study of Catalytic Residue Asp 52 in Hen Egg Lysozyme^1
- Correlation between the Differences in the Free Energy Change and conformational Energy in the Folded State of Hen Lysozymes with Gly-Pro and Pro-Gly Sequences Introduced to the Same Site
- Effect of Salt Concentration on the pK_a of Acidic Residues in Lysozyme
- Functional computed tomography imaging of tumor-induced angiogenesis : preliminary results of new tracer kinetic modeling using a computer discretization approach
- EFFECT OF CYP2C POLYMORPHISMS ON THE PHARMACOKINETICS OF PHENYTOIN IN JAPANESE PATIENTS WITH EPILEPSY
- Population Pharmacokinetics of Phenytoin in Japanese Patients with Epilepsy : Analysis with a Dose-Dependent Clearance Model
- 生産部門 無細胞蛋白質合成と非天然型アミノ酸部位特異的導入を特徴とする高難度蛋白質試料調製技術の開発 再構築型無細胞蛋白質合成系PURE system (融合発展する構造生物学とケミカルバイオロジーの最前線) -- (ターゲットタンパク研究プログラムの紹介)
- 大腸菌全蛋白質の網羅的な凝集の解析
- Simple Modeling of an Abdomen of Pregnant Women and Its Application to SAR Estimation(Electromagnetic Compatibility (EMC))
- Effect of Sucrose on Formation of the β-Amyloid Fibrils and D-Aspartic Acids in Aβ1-42
- Aggregation and Chemical Reaction in Hen Lysozyme Caused by Heating at pH 6 Are Depressed by Osmolytes, Sucrose and Trehalose^1
- Resolution and Characterization of Tryptophyl Fluorescence of Hen Egg-White Lysozyme by Quenching-and Time-Resolved Spectroscopy
- Functional and Structural Effects of Mutagenic Replacement of Asn37 at Subsite F on the Lysozyme-catalyzed Reaction
- Amino Acid Residues in Subsites E and F Responsible for the Characteristic Enzymatic Activity of Duck Egg-White Lysozyme
- Stabilization of Lysozyme against Irreversible Inactivation by Suppression of Chemical Reactions
- Effects of Additives on Irreversible Inactivation of Lysozyme at Neutral pH and 100℃
- Effect of Buffer Species on the Unfolding and the Aggregation of Humanized IgG
- A Method for the Detection of Asparagine Deamidation and Aspartate Isomerization of Proteins by MALDI/TOF-Mass Spectrometry Using Endoproteinase Asp-N
- ヌクレアーゼ型コリシンE4とE5の作用機構の研究
- コリシンE4とE5の持つヌクレアーゼ活性に関する研究 : 微生物
- リボソームに対するコリシンE3の作用 : 活性変異体を用いた再検討 : 微生物
- Analyses of Native Disulfide Bond Formations in the Early Stage of the Folding Process in Mutant Lysozymes Where the Long-Range Interactions in the Denatured State Were Modulated
- 再構築型無細胞タンパク質合成系 PURE system の開発と利用
- 2P128 転写・翻訳系がカップルしたin vitro無細胞翻訳系再構成の試み(核酸結合蛋白質,第48回日本生物物理学会年会)
- Enhancing Effect of Tumor Promoters, Phorbol Esters and Teleocidins on Nuclear Receptor-Mediated Transcription
- Identification of Protein Disulfide Isomerase as a Phorbol Ester-Binding Protein
- Pyridine-Promoted Factor-and Energy-Free Peptide Synthesis Systems Prepared from Various Organisms Including Prokaryote, Eukaryote, and Mitochondria
- Factor- and Energy-Free Peptide Synthesis Promoted by Aromatic Tertiary Amines Including Nucleic Acid-Related Compoiunds
- Template-Dependent Polypeptide Synthesis in a Factor- and Energy-Free Translation System Promoted by Pyridine
- Esterification of Eschericia coli tRNAs with D-Histidine and D-Lysine by Aminoacyl-tRNA Synthetases
- 線形動物ミトコンドリアにおけるtRNAとEF-Tuの相互作用
- プライマーとしてtRNA^/DNAを用いたHIV-1逆転写/テンプレート交換反応の効率
- ESI/Iontrap MS を用いた微量修飾ヌクレオシドの解析
- 牛ミトコンドアtRNA中に見いだされた未同定修飾塩基の構造解析
- LC/ESI MSを用いた修飾ウリジンの高感度, 特異的検出法の確立
- Trends in oncological CT imaging : clinical application of multidetector-row CT and 3D-CT imaging
- 高品質なタンパク質を作るためのPURE Technology (特集 無細胞生命科学の創成)
- 無細胞遺伝子発現システムの現状と展望
- 蛋白質合成系の完全再構成--蛋白質誕生から成熟までのピュアアプローチ (細胞における 蛋白質の一生--生成・成熟・輸送・管理・分解・病態) -- (蛋白質の生成と成熟)
- Expression and Characterization of Bovine Mitochondrial Methionyl-tRNA Transformylase
- 多義語コドンの実験的検証
- ウシミトコンドリア蛋白質合成系におけるセリルtRNA合成酵素のクローニング
- リボソームRNAの触媒機能
- リボソームとRNAワールド rRNAの触媒機能はどこまでわかってきたか
- Dynamic-Enhanced MRI Predicts Metastatic Potential of Invasive Ductal Breast Cancer
- An Analysis of Chest Wall and Diaphragm Motions in Patients With Idiopathic Scoliosis Using Dynamic Breathing MRI
- 線形動物EF-Tu相同蛋白質の大腸菌における発現
- Induction of Chromatin Damage and Distribution of Isochromatid Breaks in Human Fibroblast Cells Exposed to Heavy Ions
- Assignment of Imino Proton Signals of G-C Base Pairs and Magnesium Ion Binding: An NMR Study of Bovine Mitochondrial tRNA^_ Lacking the Entrire D Arm^1
- Substrate Specificity of tRNA (Adenine-1-)-methyltransferase from Thermus thermophilus HB27
- A Novel Screening System for Self-mRNA Targeting Proteins
- 芳香族三級アミンを用いた新規な生体外タンパク質合成系とその翻訳系の起源研究への意義
- 多義語コドンの発見--複数のアミノ酸と結合するファジ-なtRNA
- リボソーム不活性化酵素や抗生物質を用いたピリジン蛋白質合成系の解析
- 大腸菌23SリボソームRNAによるペプチド結合形成反応
- ピリジン存在下の蛋白質合成系におけるリボソーム構成要素の解析
- Insect Lysozyme from House Fly (Musca domestica) Larvae: Possible Digestive Function Based on Sequence and Enzymatic Properties
- ウシミトコンドリアの翻訳開始機構におけるMet-tRNA transformylase(MTF)
- Extraskeletal mesenchymal chondrosarcoma of the cervical meninx
- Modification Defect at Anticodon Wobble Nucleotide of Mitochondrial tRNAs^(UUR) with Pathogenic Mutations of Mitochondrial Myopathy, Encephalopathy, Lactic Acidosis, and Stroke-like Episodes
- 蛋白質合成系におけるリボソームRNAの機能
- タンパク質合成系を利用したアミノ酸の鋳型重合
- 生体外蛋白質合成系の研究・開発と利用 有機触媒を用いた新規合成系に期待
- 哺乳類ミトコンドリアにおける新規翻訳因子の発見
- Kinetic Measurement of the Interaction between a Lysozyme and Its Immobilized Substrate Analogue by Means of Surface Plasmon Resonance
- Analyses of Native Disulfide Bond Formations in the Early Stage of the Folding Process in Mutant Lysozymes Where the Long-Range Interactions in the Denatured State Were Modulated
- Magnetic resonance imaging of hepatocellular carcinoma : a pictorial review of novel insights into pathophysiological features revealed by magnetic resonance imaging
- 無細胞合成系PURE systemを基盤とした応用と人工細胞の構築
- A pictorial review of benign hepatocellular nodular lesions : comprehensive radiological assessment incorporating the concept of anomalous portal tract syndrome
- 1S4p06 東京大学iGEM学生チームiGEM2011プロジェクトの紹介(iGEMを通した合成生物学研究者の育成,ワークショップ)
- Construction of a Screening System for Selecting Lysozyme Mutants Unable to Form a Stable Structure from Random Mutants
- tRNAの翻訳機能と修飾塩基(蛋白質合成におけるRNAの機能) (核酸化学の新展開--新しい機能性分子を求めて) -- (核酸の機能)
- 大腸菌全タンパク質の凝集性とシャペロン効果の網羅的な解析
- Influence of Mutations of the N-Cap Residue, Gly4, on Stability and Structure of Hen Lysozyme.
- A Mutation Study of Catalytic Residue Asp 52 in Hen Egg Lysozyme.
- An Improved Method for Preparing Lysozyme with Chemically 13C-Enriched Methionine Residues Using 2-Aminothiophenol as a Reagent of Thiolysis.
- Detection of a Local Interaction of Hen Lysozyme under Highly Denaturing Conditions Using Chemically 13C-Enriched Methionine Resonance.