1999 Fiscal Year Annual Research Report

ラン藻由来の新規なプラストシアニンのX線構造解析および分子進化に関する研究

Research Project

Project/Area Number	11780495
Research Institution	Osaka University
Principal Investigator	井上豪大阪大学, 大学院・工学研究科, 講師 (20263204)
Keywords	ラン藻 / Synechococcus sp.PCC 7942 / プラストシアニン / X線構造解析 / 酸化体 / 還元体 / 分子進化 / 電子伝達 / 光合成
Research Abstract	ラン藻Synechococcus sp.PCC7942由来の光合成電子伝達蛋白質であるプラストシアニン(PC)の酸化体および還元体のX線構造解析をそれぞれ1.9Å分解能、1.8Å分解能で行った。通常PCは、高等植物では99残基、緑藻類では97残基で、そのアミノ酸総数が保存されている。平成11年度に論文報告を行ったシダ植物由来のPCでは102残基で、これまで発見されている高等植物由来のPC中では最長であったが、本研究課題で取り組んだPCは、その逆で、これまで発見された中では最も短い91残基で構成される。蛋白質レベルでは発見されず、その存在は遺伝子から見つけられた。もう1つの特徴として、電子伝達のパ-トナーの認識部位と考えられている酸性を帯びた結合部位の構造を持たない。本研究では、新規なラン藻由来PCを大腸菌を使って大量発現させ、結晶化と構造解析を行ったものである。解析の結果、アミノ酸の相同性が極めて低いにも関わらず、全体構造は類似していた。電子伝達の入り口と考えられているTyr83の芳香環の上にAgr88が覆い被さり、π-πスタッキング相互作用が見られた。pH5.0で結晶が得られたが、その酸化体と還元体のX線構造の平均の構造のずれが0.08Åと小さいにも関わらず、His配位子が0.36Åもずれたり、銅イオンから約12A離れたArg88が0.42Åずれていた。銅から遠隔の場所で微細ではあるが有意な構造変化が得られ、電子伝達の窓口であるTyr83の役割が重要であり、分子進化によって失われることなく

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] T.Inoue et al.: "Crystallization and preliminary X-ray Study of iso-2 Azurin from Methylotrophic Bacterium, Methylomnas J."Acta Cryst.,. D55,. 307-309 (1999)
[Publications] N.Shibata et al.: "Novel Insight into the Copper-Ligands Geometry in the Crystal Structure of Ulva Pertusa Plastocyanin at 1.6 A Resolution"J. Biol, Chem.,. 274,. 4225-4230 (1999)
[Publications] T. Inoue, et al.: "Crystallization and Preliminary X-ray Analysis of Plastocyanin from Cyanobacterum Synechococcus sp. PCC 7942"Acta. Cryst.,. D55,. 683-684 (1999)
[Publications] T. Inoue, et al.: "Crystal Structure Determinations of Oxidized and Reduced Plastocyanin from the Cyanobacterium Synechococcus sp. PCC 7942"Biochemistry,. 38,. 6063-6069 (1999)
[Publications] T.Kohzuma et al.: "The Structure and Unusual pH-dependence of Pastocyanin from Fern Drypeteris crassirhizoma: the Protonation of an Active-Site Histidine is Hindered by p-p interactions"J. Biol. Chem.,. 274,. 11817-11823 (1999)
[Publications] Y.Kai et al.: "Three-Dimensional Structure of Phosphoenolphruvate Carboxylase: A Proposed Novel Mechanism for Allosteric Inhibition"Proc. Natl. Acad. Sci. USA,. 96,. 823-828 (1999)
[Publications] H.Sugawara et al.: "Cryatal Structure of Wild-type and Mutant Plastocyanins from Higher Plant Silene"J. Biochem.,. 125,. 899-903 (1999)
[Publications] T.Inoue et al.: "Crystal Structure Detemination of Oxidized and Reduced Pseudoazurin from Achromobacter Cycloclaters. Concerted Movement of Copper Site in Redox Forms with the Rerrangement of Hydrogen Bond at a Remote Histidine,"J,Biol,Chem.,. 274,. 17845-17852 (1999)
[Publications] T.Inoue et al.: "Structure Comparison between Oxidized and Reduced Plastocyanin from a Fern, Drypteris crassirhizoma"Biochemistry,. 38,. 13853-13861 (1999)
[Publications] H.Matsumura et al.: "Crystallization and preliminary X-ray diffraction studies of C4-form phosphoenolpyruvate carboxylase from maize"Acta Cryst.,. 55,. 1937-1938 (1999)
[Publications] S.Suzuki et al.: "Structure-function relationships of copper-containing nitrite reductases"Coordination Chemistry Reviews,. 192,. 245-265 (1999)
[Publications] H.Hashimoto et al.: "Hyperthemostable protein structure maintained by intra and inter-helix ion-pairs archaeal O-6-methylguanine-DNA methyltransferase"J. Mol. Biol.. 292,. 707-716 (1999)
[Publications] S.Suzuki et al.: "Spectroscopic and electrochemical properties of two azurins (Az-iso 1and Az-iso 2) from the Obligate methylotroph Methylomonas sp. Strain J and structure of novel Az-iso 2"J. Biol, lnorg, Chem.,. 4,. 749-758 (1999)
[Publications] H.Sugawara et al.: "Cryatal Structure of Carboxylase reaction-oriented ribulose 1, 5-bisphosphate carboxylase/oxygenase from a thermophilic red alga, Caldieria partita"J. Biol. Chem,. 274,. 15655-15661 (1999)
[Publications] H.Hishimoto et al.: "Crystallographic Studies on a Family B DNA Polymerase from Hyperthermophilic Archaeon Pyrococcus kodakaraensis Strain KOD1"J. Biochem.,. 125,. 983-986 (1999)
[Publications] H.Matsumura et al.: "Plausible Phosphoenolpyruvate binding site revealed by 2.6 A structure of Mn2+bound phosphoenolpruvate carboxylase from Escherichia coli"FEBS LETT.,. 458,. 93-96 (1999)

1999 Fiscal Year Annual Research Report

ラン藻由来の新規なプラストシアニンのX線構造解析および分子進化に関する研究

Principal Investigator

井上 豪 大阪大学, 大学院・工学研究科, 講師 (20263204)

Research Products

[Publications] T.Inoue et al.: "Crystallization and preliminary X-ray Study of iso-2 Azurin from Methylotrophic Bacterium, Methylomnas J."Acta Cryst.,. D55,. 307-309 (1999)

[Publications] N.Shibata et al.: "Novel Insight into the Copper-Ligands Geometry in the Crystal Structure of Ulva Pertusa Plastocyanin at 1.6 A Resolution"J. Biol, Chem.,. 274,. 4225-4230 (1999)

[Publications] T. Inoue, et al.: "Crystallization and Preliminary X-ray Analysis of Plastocyanin from Cyanobacterum Synechococcus sp. PCC 7942"Acta. Cryst.,. D55,. 683-684 (1999)

[Publications] T. Inoue, et al.: "Crystal Structure Determinations of Oxidized and Reduced Plastocyanin from the Cyanobacterium Synechococcus sp. PCC 7942"Biochemistry,. 38,. 6063-6069 (1999)

[Publications] T.Kohzuma et al.: "The Structure and Unusual pH-dependence of Pastocyanin from Fern Drypeteris crassirhizoma: the Protonation of an Active-Site Histidine is Hindered by p-p interactions"J. Biol. Chem.,. 274,. 11817-11823 (1999)

[Publications] Y.Kai et al.: "Three-Dimensional Structure of Phosphoenolphruvate Carboxylase: A Proposed Novel Mechanism for Allosteric Inhibition"Proc. Natl. Acad. Sci. USA,. 96,. 823-828 (1999)

[Publications] H.Sugawara et al.: "Cryatal Structure of Wild-type and Mutant Plastocyanins from Higher Plant Silene"J. Biochem.,. 125,. 899-903 (1999)

[Publications] T.Inoue et al.: "Crystal Structure Detemination of Oxidized and Reduced Pseudoazurin from Achromobacter Cycloclaters. Concerted Movement of Copper Site in Redox Forms with the Rerrangement of Hydrogen Bond at a Remote Histidine,"J,Biol,Chem.,. 274,. 17845-17852 (1999)

[Publications] T.Inoue et al.: "Structure Comparison between Oxidized and Reduced Plastocyanin from a Fern, Drypteris crassirhizoma"Biochemistry,. 38,. 13853-13861 (1999)

[Publications] H.Matsumura et al.: "Crystallization and preliminary X-ray diffraction studies of C4-form phosphoenolpyruvate carboxylase from maize"Acta Cryst.,. 55,. 1937-1938 (1999)

[Publications] S.Suzuki et al.: "Structure-function relationships of copper-containing nitrite reductases"Coordination Chemistry Reviews,. 192,. 245-265 (1999)

[Publications] H.Hashimoto et al.: "Hyperthemostable protein structure maintained by intra and inter-helix ion-pairs archaeal O-6-methylguanine-DNA methyltransferase"J. Mol. Biol.. 292,. 707-716 (1999)

[Publications] S.Suzuki et al.: "Spectroscopic and electrochemical properties of two azurins (Az-iso 1and Az-iso 2) from the Obligate methylotroph Methylomonas sp. Strain J and structure of novel Az-iso 2"J. Biol, lnorg, Chem.,. 4,. 749-758 (1999)

[Publications] H.Sugawara et al.: "Cryatal Structure of Carboxylase reaction-oriented ribulose 1, 5-bisphosphate carboxylase/oxygenase from a thermophilic red alga, Caldieria partita"J. Biol. Chem,. 274,. 15655-15661 (1999)

[Publications] H.Hishimoto et al.: "Crystallographic Studies on a Family B DNA Polymerase from Hyperthermophilic Archaeon Pyrococcus kodakaraensis Strain KOD1"J. Biochem.,. 125,. 983-986 (1999)

[Publications] H.Matsumura et al.: "Plausible Phosphoenolpyruvate binding site revealed by 2.6 A structure of Mn2+bound phosphoenolpruvate carboxylase from Escherichia coli"FEBS LETT.,. 458,. 93-96 (1999)

井上豪大阪大学, 大学院・工学研究科, 講師 (20263204)