Molecular Cloning and Functional Expression of cDNA Encoding the Cysteine Proteinase Inhibitor Sca from Sunflower Seeds

スポンサーリンク

概要

• 論文の詳細を見る

• Sunflower cystatin a (Sca) is distinguished from other phytocystatins by its lack of the N-terminal about 20 amino acids, resulting in the absence of the evolutionarily conserved Gly residue. The cDNA encoding Sca was amplified by PCR methods. The cDNA consists of 520 nucleotides and includes an open reading frame encoding a polypeptide of 98 amino acids. Comparison of the deduced amino acid sequence with the Sca protein sequence indicated that the deduced sequence has an extra 15 amino acids and one amino acid at the N-and C-termini, respectively. This result suggests that Sca is synthesized as a preprotein (preSca) and proteolytic cleavages at peptide bonds may give rise to the mature Sca. To address this assumption and also to investigate the significance of the N-terminal extension sequence to Sca for inhibitory activity, a recombinant pre-Sca (rpre-Sca), in which the N-terminal extension was fused to the matured Sca, and a recombinant matured Sca (rSca) were overproduced in Escherichia coli cells. Incubation of the rpre-Sca with a seed extract resulted in a mobility by SDS-PAGE that was the same as rSca, demonstrating a proteolytic cleavage by endogenous proteinases. The rSca and rpre-Sca proteins were further characterized with respect to inhibitory activity and sensorgrams of the interaction with papain. The result showed that rpre-Sca had stronger inhibitory activity than rSca, and that the increased activity toward papain was due to a lower dissociation rate constant. This finding indicates that the N-terminal region of rpre-Sca increases the inhibitory activity by stabilizing the rpre-Sca and papain complex.

• 社団法人日本農芸化学会の論文
• 2001-04-23

著者

• YAMASAKI Nobuyuki

  Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University

• KIMURA Makoto

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

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Faculty of Agriculture, Graduate School, Kyushu University

• Kimura M

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• INANAGA Hideko

  Laboratory of Biochemisty, Graduate School, Kyushu University

• TSUKIGATA Keiko

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad

• DOI-KAWANO Keiko

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad

• Doi-kawano Keiko

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

• 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

• Inanaga Hideko

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

• Yamasaki N

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• Kouzuma Yoshiaki

  Laboratory Of Food Molecular Functionality College Of Agriculture Ibaraki University

• Kouzuma Yoshiaki

  Laboratory Of Food Functionality School Of Agriculture Ibaraki University

• Kawabe Masato

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

• Tsukigata Keiko

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

• 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

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University

• 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)
• 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
• Biotransformation of the Mycotoxin, Zearalenone, to a Non-estrogenic Compound by a Fungal Strain of Clonostachys sp.(Organic Chemistry)
• Structural Evidence for Endocrine Disruptor Bisphenol A Binding to Human Nuclear Receptor ERRγ
• Cloning of the Gene Encoding DNA Binding Protein HU from Bacillus stearothermophilus and Its Expression in Escherichia coli
• The Evolution of the Phenylacetic Acid Degradation Pathway in Bacteria
• 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)
• Vertical changes in bacterial and archaeal communities with soil depth in Japanese paddy fields(Soil Biology)
• Molecular characterization of methane-oxidizing bacteria associated with rice straw decomposition in a rice field(Soil 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
• The Action of Rice Branching Enzyme I (BEI) on Starches
• 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)
• 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
• Suppression of Ralstonia solanacearum in Soil Following Colonization by Other Strains of R. solanacearum
• Morphology, host range and phylogenetic diversity of Sphingomonas phages in the floodwater of a Japanese paddy field(Soil Biology)
• Abundance of bacteriophages of common heterotrophic bacteria in the floodwater of a Japanese paddy field(Soil Biology)
• Seasonal variations in the abundance of virus-like particles and bacteria in the floodwater of a Japanese paddy field(Soil Biology)
• Requirement for C-terminal Extension to the RNA Binding Domain for Efficient RNA Binding by Ribosomal Protein L2(Biochemistry & Molecular Biology)
• 5-8 Distribution of Photosynthetically Fixed C in Irrigated Rice : I. Contribution to Soil Dissolved Organic C and Microbial Biomass
• Effect of Rice Straw Application on CH_4 Emission from Paddy Fields : IV. Influence of Rice Straw Incorporated during the Previous Cropping Period
• Effect of Rice Straw Application on CH_4 Emission from Paddy Fields : III. Effect of Incorporation Site of Rice Straw on CH_4 Emission Rates and Their Variation among Shoots of a Rice Plant
• Methane Production and Its Fate in Paddy Fields : V. Fate of Methane Remaining in Paddy Soil at Harvesting Stage
• Activities of Soil Enzymes in Different Land-Use Systems in Middle Terrace Areas of Lampung Province, South Sumatra, Indonesia
• The Contribution of Peripheral Stem-Loops to the Catalytic Activity of Archaeal RNase P RNA from Pyrococcus horikoshii OT3
• Contribution of Gln9 and Phe8O to Substrate Binding in Ribonuclease MC1 from Bitter Gourd Seeds
• Amino Acids Conserved at the C-Terminal Half of the Ribonuclease T2 Family Contribute to Protein Stability of the Enzymes
• Expression and Mutational Analysis of Amino Acid Residues Involved in Catalytic Activity in a Ribonuclease MC1 from the Seeds of Bitter Gourd
• Molecular analysis of the ammonia oxidizing bacterial community in the surface soil layer of a Japanese paddy field(Soil Biology)
• 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)
• Identification of Nucleotide Residues Essential for RNase P Activity from the Hyperthermophilic Archaeon Pyrococcus horikoshii OT3
• 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)
• 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)
• A Novel Adsorbent for Affinity Chromatography of Lysozyme

もっと見る 閉じる

スポンサーリンク