Expression and Mutational Analysis of Amino Acid Residues Involved in Catalytic Activity in a Ribonuclease MC1 from the Seeds of Bitter Gourd

スポンサーリンク

概要

• 論文の詳細を見る

• The ribonuclease MC1 (RNase MC1) from seeds of bitter gourd (Momordica charantia) consists of 190 amino acids and belongs to the RNase T2 family, including fungal RNase typified by RNase Rh from Rhizopus niveus. We expressed RNase MC1 in Escherichia coli cells and made use of site-directed mutagenesis to identify essential amino acid residues for catalytic activity. Mutations of His34 and His88 to Ala completely abolished the enzymatic activity, and considerable decreases in the enzymatic activity were observed in cases of mutations of His83,Glu84,and Lys87,when yeast RNA was used as a substrate. Kinetic parameters for the enzymatic activity of the mutants of His83,Glu84,and Lys87 were analyzed using a dinucleoside monophosphate CpU. K_m values for the mutants were approximately like that for wild-type, while k_<cat> values were decreased by about 6 to 25-fold. These results suggest that His34,His83,Glu84,Lys87,and His88 in RNase MC1 may be involved in the catalytic function. These observation suggests that RNase MC1 from a plant catalyzes RNA degradation in a similar manner to that of fungal RNases.

• 社団法人日本農芸化学会の論文
• 2000-03-23

著者

• YAMASAKI Nobuyuki

  Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University

• FUNATSU Gunki

  Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University

• KIMURA Makoto

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• Kimura M

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• Yamasaki Nobuyuki

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• Yamasaki Nobuyuki

  Laboratory Of Biochemistry And Laboratory Of Protein Chemistry And Engineering Faculty Of Agricultur

• Yamasaki N

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• Kawabe Masato

  United Graduate School Of Agricultural Science Tokyo University Of Agriculture And Technology (tuat)

• Hino M

  Kyushu Univ. Fukuoka Jpn

• Funatsu G

  Kyushu Univ. Fukuoka Jpn

• Funatsu Gunki

  Laboratory Of Biochemistry And ** Laboratory Of Protein Chemistry & Engineering Faculty Of Agric

• Numata T

  Joetsu Univ. Education Niigata Jpn

• Numata Tomoyuki

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• KASHIBA Tohru

  Laboratory of Protein Chemistry and Engineering, Graduate School of Genetic Resources Technology, Ky

• HINO Madoka

  Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University

• ISHIGURO Masatune

  Laboratory of Protein Chemistry and Engineering, Graduate School of Genetic Resources Technology, Ky

• Hino Madoka

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Kashiba Tohru

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Genetic Resources Technology Kyus

• Ishiguro Masatune

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Genetic Resources Technology Kyus

• Yamasaki Nobuyuki

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Graduate School Faculty Of Agr

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Graduate School Faculty Of Agr

• Yamasaki Nobuyuki

  Laboratory Of Agricultural Process Engineering Faculty Of Agriculture Kyushu University

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Agricultural Chemistry Faculty And Graduate School Of Agric

• KIMURA Makoto

  Laboratory for Remediation Research, Plant Science Center, RIKEN

関連論文

• Archaeal Homologs of Human RNase P Protein Pairs Pop5 with Rpp30 and Rpp21 with Rpp29 Work on Distinct Functional Domains of the RNA Subunit
• Some Properties of Ricin D Modified with a Methoxypolyethylene Glycol Derivative(Biological Chemistry)
• Further Studies on the Histidine Residue Located at the High Affinity Saccharide-binding Site of Ricin E(Biological Chemistry)
• Identification of the Tryptophan Residue Present in the Low Affinity Saccharide-binding Site of Ricin E(Biological Chemistry)
• Significance of the Histidine Residue at the High Affinity Saccharide-binding Site of Ricin E(Biological Chemistry)
• Significance of the Tryptophan Residue at the Low Affinity Saccharide-binding Site of Ricin E(Biological Chemistry)
• Chemical Modification of Ricin D with N-Acetylimidazole(Biological Chemistry)
• The Interaction of Castor Bean Lectins with Galactomannan(Biological Chemistry)
• Denaturation of Ricin D in Frozen Aqueous Solutions(Biological Chemistry)
• Base Specificity of Two Plant Seed Ribonucleases from Momoridica charantia and Luffa cylindrica
• Crystal Structure of hyperthermophilic Archaeal Initiation Factor 5A : A Homologue of Eukaryotic Initiation Factor 5A (eIF-5A)
• The Structure of the Archaebacterial Ribosomal Protein S7 and Its Possible Interaction with 16S rRNA^1
• Resolution and Characterization of Tryptophyl Fluorescence of Hen Egg-White Lysozyme by Quenching-and Time-Resolved Spectroscopy
• Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata
• Expression in Cereal Plants of Genes That Inactivate Fusarium Mycotoxins(Biochemistry & Molecular Biology)
• Proteome Analysis of Wheat Lemma(Biochemistry & Molecular Biology)
• Studies on the Carbohydrate Binding Sites of the Hemolytic Lectin CEL-III Isolated from the Marine Invertebrate Cucumaria echinata
• Cytotoxicity Induced by Erythrina variegata Serine Proteinase Inhibitors in Tumor Hematopoietic Stem Cell Lines
• Methanogenic archaeal communities developed in paddy fields in the Kojima Bay polder, estimated by denaturing gradient gel electrophoresis, real-time PCR and sequencing analyses(Soil Biology)
• Structural Changes in Ribonuclease P RNA in the Hyperthermophilic Archaeon Pyrococcus horikoshii OT3 Induced on Interaction with Proteins
• Crystal Structure and Functional Analysis of an Archaeal Chromatin Protein Alba from the Hyperthermophilic Archaeon Pyrococcus horikoshii OT3
• Origin and Properties of Humus in the Subsoil of Irrigated Rice Paddles : IV. Effect of Water Percolation on the Binding Type of Humus in a Submerged Plow Layer Soil
• Comparison of Humus Composition in the Subsoil of Japanese Paddy and Upland Fields
• Origin and Properties of Humus in the Subsoil of Irrigated Rice Paddies : III. Changes in Binding Type of Humus in Submerged Plow Layer Soil
• Origin and Properties of Humus in the Subsoil of Irrigated Rice Paddies : II. Effects of Subsoil Constituents and Drainage on the Accumulation of Organic Matter Leached from the Plow Layer Soil
• Origin and Properties of Humus in the Subsoil of Irrigated Rice Paddies : I. Leaching of Organic Matter from Plow Layer Soil and Accumulation in Subsoil
• Genomic Cloning of Ribonucleases in Nicotiana glutinosa Leaves, as Induced in Response to Wounding or to TMV-Infection, and Characterization of Their Promoters(Biochemistry & Molecular Biology)
• Spectroscopic Analysis of the Cytoagglutinating Activity of Abrin-b Isolated from Abrus precatorius Seeds against Leukemic Cells
• Contribution of Conserved Asn Residues to the Inhibitory Activities of Kunitz-Type Protease Inhibitors from Plants
• Protein Engineering of Novel Proteinase Inhibitors and Their Effects on the Growth of Spodoptera exigua Larvae(Biochemistry & Molecular Biology)
• Calcium Ions Stabilize a Protein Structure of Hemolytic Lectin CEL-III from Marine Invertebrate Cucumaria echinata
• Molecular Cloning and Functional Expression of cDNA Encoding the Cysteine Proteinase Inhibitor Sca from Sunflower Seeds
• Conformation of the Primary Binding Loop Folded through an Itramolecular Iteraction Contributes to the Strong Chymotrpsin Inhibitory Activity of the Chymotrypsin Inhibitor from Erythrina variegata Seeds
• Molecular Cloning, Functional Expression, and Mutagenesis of cDNA Encoding a Cysteine Proteinase Inhibitor from Sunflower Seeds
• Chymotrypsin Inhibitor from Erythrina variegata Seeds : Involvement of Amino Acid Residues within the Primary Binding Loop in Potent Inhibitory Activity toward Chymotrypsin
• Cloning, Expression, and Mutagenesis of Trypsin Inhibitor ETIb from Erythrina variegata Seeds
• The Tissue-Type Plasminogen Acitvator Inhibitor ETIa from Erythrina variegata: Structural Basis for the Inhibitory Activity by Cloning, Expression, and Mutagenesis of the cDNA Encoding ETIa
• Molecular Cloning of cDNA That Encodes Chymotrypsin Inhibitor ECI from Erythrina variegata Seeds and Its Expression in Escherichia coli
• Inhibitory Potency of Erythrina variegata Proteinase Inhibitors toward Serine Proteinases in the Blood Coagulation and Fibrinolytic Systems
• Purification, Characterization, and Sequencing of Two Cysteine Proteinase Inhibitors, Sca and Scb, from Sunflower (Helianthus annuus) Seeds
• Amino Acid Sequence of Chymotrypsin Inhibitor ECI from the Seeds of Erythrina variegata (LINN.) var. Orientalis
• Isolation and Primary Structure of Proteinase Inhibitors from Erythrina variegata (LINN.) var. Orientalis Seeds
• On the Interaction of Ribosomal Protein L5 with 5S rRNA(Biochemistry & Molecular Biology)
• Arginine-55 in the β-Arm is Essential for the Activity of DNA-Binding Protein HU from Bacillus stearothermophilus
• Contribution of a Salt Bridge to the Thermostability of DNA Binding Proten HU from Bacillus stearothermophilus Determined by Site-Directed Mutagenesis
• Characterization of the Archaeal Ribonuclease P Proteins from Pyrococcus horikoshii OT3
• Effect of elevated atmospheric CO_2 concentration on ammonia oxidizing bacteria communities inhabiting in rice roots(Soil Biology)
• A Wheat Xylanase Inhibitor Gene, Xip-I, but Not Taxi-I, Is Significantly Induced by Biotic and Abiotic Signals That Trigger Plant Defense
• New TAXI-type Xylanase Inhibitor Genes are Inducible by Pathogens and Wounding in Hexaploid Wheat
• Brefeldin A Protects Ricin-Induced Cytotoxicity in Human Cancer KB Cell Line, but not in its Resistant Counterpart with Altered Golgi Structures
• Cloning of the Gene Encoding DNA Binding Protein HU from Bacillus stearothermophilus and Its Expression in Escherichia coli
• Amino Acid Sequence and Carbohydrate-binding Analysis of the N-acetyl-D-galactosamine-specific C-Type Lectin, CEL-I, from the Holothuroidea, Cucumaria echinata(Biochemistry & Molecular Biology)
• Primary Structure of 6.5k-Arginine/Glutamate-rich Polypeptide from the Seeds of Sponge Gourd (Luffa cylindrica)
• Chemical Modification of the Hemolytic Lectin CEL-III by Succinic Anhydride : Involvement of Amino Groups in the Oligomerization Process
• Chemical Modifications of Momordin-a and Luffin-a, Ribosome-Inactivating Proteins from the Seeds of Momordica charantia and Luffa cylindrica : Involvement of His140,Tyr165,and Lys231 in the Protein-Synthesis Inhibitory Activity
• Purification and Characterization of Two Lectins from the Sea Cucumber Stichopus japonicus
• An Assay for Lectin Activity Using Dextran Derivatives
• Molecular Mechanism for Pore-formation in Lipid Membranes by the Hemolytic Lectin CEL-III from Marine Invertebrate Cucumaria echinata
• A Novel Amino Acid Derivative for Fluorescence-labeling of Carboxyl Groups in Proteins(Biological Chemistry)
• Amino Acid Sequences of the Two Smallest Trypsin Inhibitors from Sponge Gourd Seeds(Biological Chemistry)
• Temperature- and pH-Dependent Cytotoxic Effect of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata on Various Cell Lines
• Carbohydrate-Binding Properties of the Hemolytic Lectin CEL-III from the Holothuroidea Cucumaria echinata as Analyzed Using Carbohydrate-Coated Microplate
• Amino Acid Sequence of a C-Type Lectin CEL-IV from the Marine Invertebrate Cucumaria echinata
• Improvement of the Thermostability of Pyruvate Decarboxylase by Modification with an Amylose Derivative
• Isolation and Amino Acid Sequence of a Protein-synthesis Inhibitor from the Seeds of Rye (Secale cereale)
• Purification and Characterization of Three Mitogenic Lectins from the Roots of Pokeweed : (Phytolacca americana)
• Complete Amino Acid Sequence of Luffin-a, a Ribosome-inactivating Protein from the Seeds of Luff a cylindrica(Biological Chemistry)
• Complete Amino Acid Sequence of Luffin-b, a Ribosome-Inactivating Protein from Sponge Gourd (Luffa cylindrica) Seeds(Biological Chemistry)
• Structural Analysis of N-Linked Oligosaccharide of Mitogenic Lectin-B from the Roots of Pokeweed (Phytolacca americana)
• Purification of Chymotrypsin Inhibitors from Larval Hemolymph of the Silkworm, Bombyx mori
• Expression and Mutational Analysis of Amino Acid Residues Involved in Catalytic Activity in a Ribonuclease MC1 from the Seeds of Bitter Gourd
• The Amino Acid Sequence of Mitogenic Lectin-B from the Roots of Pokeweed : Phytolacca americana
• Amino Acid Sequence and Some Properties of Lectin-D from the Roots of Pokeweed (Phytolacca americana)
• The Complete Amino Acid Sequence of Lectin-C from the Roots of Pokeweed (Phytolacca americana)
• Role of the Histidine Residue at the Saccharide-binding Site of Castor Bean Hemagglutinin(Biological Chemistry)
• Chemical Modification of Castor Bean Hemagglutinin with Diethylpyrocarbonate(Biological Chemistry)
• States and Roles of Tryptophan Residues in Abrus precatorius Agglutinin(Biological Chemistry)
• Identification of the Tryptophan Residue Located at the Saccharide Binding Site of Castor Bean Hemagglutinin(Biological Chemistry)
• Purification and Characterization of Abrus Precatorius agglutinin
• Binding of Saccharides to Abrin-b Isolated from Abrus precatorius Seeds
• Chemical Modification of Tryptophan Residues in Abrin-a(Biological Chemistry)
• The Interaction of Abrin-a with Saccharides as Analyzed by Equilibrium Dialysis and Spectroscopy(Biological Chemistry)
• Spectroscopic Analysis of the Interaction of Ricin D with Urea(Biological Chemistry)
• Properties of Cuticular Phenoloxidase from Pupae of the Housefly, Musca domestica L.(Biological Chemistry)
• On the Mechanism of Lysozyme Catalysis I. Hydrolysis of 1-Benzoyl 2-Acetamido-2-Deoxy-β-D-Glucopyranoside
• Hydrophobicities of Ricin D and Its Constituent Polypeptide Chains(Biological Chemistry)
• Lysyl-Aspartic Acid Derivatives as Model Compounds of Growth Hormone to Induce Lipolysis(Biological Chemistry)
• Complete Amino Acid Sequence of Luffin-a, a Ribosome-inactivating Protein from the Seeds of Sponge Gourd (Luffa cylindrica)(Biological Chemistry)
• Isolation and Characterization of Two Momordins, Ribosome-Inactivating Proteins from the Seeds of Bitter Gourd (Momordica charantia)
• Positions of Disulfide Bonds in Rye (Secale cereale) Seed Chitinase-a
• Complete Amino Acid Sequence of Chitinase-A from Leaves of Pokeweed (Phytolacca americana)
• The Complete Amino Acid Sequence of Chitinase-B from the Leaves of Pokeweed (Phytolacca americana)
• Primary Structures of N-Linked Oligosaccharides of Momordin-a, a Ribosome-inactivating Protein from Momordica charantia Seeds(Biological Chemistry)
• Fluorescence Spectroscopic Properties of an Active Lysozyme Derivative : Kyn 62-Lysozyme(Biological Chemistry)
• Preparation and Immunological Characterization of β-Lactoglobulin-Amylose Conjugate
• Reduction of the Immunogenicity of β-Lactoglobulin from Cow's Milk by Conjugation with a Dextran Derivative
• Ultraviolet Difference Spectroscopic Analysis of the Binding of a Substrate Analogue to an Active Lysozyme Derivative: Kyn 62-Lysozyme (Biological Chemistry)
• Chemical Modification of Tryptophan Residues in Castor Bean Hemagglutinin
• A Novel Adsorbent for Affinity Chromatography of Lysozyme

もっと見る 閉じる

スポンサーリンク