The Nobel Prize in Chemistry 1997

The Na+/K+-ATPase ion pump history
1950's - British researchers Richard Keynes and Alan Hodgkin note: -that when a nerve is stimulated, Na+ flows into the nerve cell. - that Na+ is probably transported out of the cell when it is restored and ATP is required. - that Na+ transport from the cell can be inhibited by inhibiting ATP synthesis. 1957 - Jens C. Skou finds an ATPase (Na+/K+-ATPase ) that is activated by sodium and potassium ions. He was the first to describe an ion pump - an enzyme that gives directed transport of ions through a cell membrane: a fundamental mechanism in every living cell. 1961 Other ion pumps requiring ATP are discovered.
ATP的合成
主要地点： 线粒体呼吸作用及 植物叶绿体光合作用
线粒体
叶绿体
驱动力： 膜内外两侧的H+浓度梯度 催化反应的酶： ATP合成酶(ATP synthase)
ATP synthase is found in chloroplast(叶绿体) and mitochondrial membranes (线粒体 膜) and in the cytoplasmic membrane of bacteria(细菌细胞膜).
＋ATP
ATP synthase history
1937 - Herman Kalckar (Denmark) establishes that ATP synthase is linked with cell respiration. 1961 - The American Ephraim Racker isolates the F1 part of the ATP synthase. 1961 – Peter Mitchell (UK) shows that cell respiration leads to differing concentrations of hydrogen ions (pH) inside and outside the mitochondrial membrane(细胞呼吸作用使线粒体膜内外两侧产生H+浓度梯度). 1964 - Paul D. Boyer proposes that ATP is synthesized through structural changes in the ATP synthase enzyme. 1973 - Boyer ("The Binding Change Mechanism").
1981 - John E. Walker determines the DNA sequence of the genes encoding the proteins in ATP synthase. 1994 - The structure of the F1 part of the ATP synthase is determined by Walker and co-workers. 1996-1997 - The hypothesis that parts of ATP synthase rotate during the synthesis and hydrolysis of ATP is demonstrated chemically (Richard Cross, USA), spectroscopically (Wolfgang Junge, Germany) and microscopically (Masasuke Yoshida, Japan).
A difference in hydrogen ion concentration across the membrane drives the enzyme to synthesise ATP.
F1
Boyer: ATP synthase as a waterdriven hammer minting coins. F0 part is the wheel, flow of H+ is the waterfall, the structural changes in F1 lead to 3 coins in the ATP currency being minted for each turn of the wheel.
ATP synthase is a molecular machine that work like a turbine（涡轮机）to convert the energy stored in a proton gradient into mechanical, rotational energy, and then into chemical energy stored in the bond energy of ATP.
F1
F0
c亚基在膜上形成一个可旋转的环 a亚基有跨膜运输的H+通道
c ring of a chloroplast ATP synthase.(2000)
Walker 1994 获得 结晶体，发表F1 头部 from cows 详细原子模型， 提供重要的结构 学证据。 F1头部：每个F1 含有3个ATP催化位点，每个亚基各一个。 中央亚基为轴，从F0膜延伸到F1头部，其顶端高度不对称， 任何瞬间其不同部分与3个亚基接触，使每个亚基产生3种不 同构象(O/L/T)，只有每个亚基旋转到与处于某一特定构象(T) 1994 时才能催化ADP合成ATP。 每一个催化循环中， 亚 基旋转360º ，3个亚基都 能依次经过3种不同的构 象变化。
Jens C. Skou Aarhus University Denmark
Life's energy currency, ATP 生命的活动和维持需要消
耗能量，生物体本身不能创 造新的能量，只能依赖外 部能量的输入。
几乎所有地球生命所需的 能量都来自太阳。
Originally, the energy comes from the sun.
The Nobel Prize in Chemistry 1997
½ prize to Jens C. Skou, for “the first discovery of an ion-transporting enzyme, Na+/K+-ATPase.”
•动物细胞的Na+/K+ 泵( Na+/K+-ATPase)
ATP不到1g, 即每个细胞每秒钟大约可形成一千万个ATP，同时 有同样量的ATP被水解，产生能量供生命活动。
以ATP形式储存的能量在各类生命合成代谢中起递能作用。
Every day an adult converts a quantity of ATP corresponding to at least half his or her body weight, and during hard work the quantity can rise to almost a ton(吨).
(ADP). 标准状态下，每摩尔ATP水解为ADP时，可产生30.5 能量。
kJ的
萤火虫利用ATP提供能量发出荧光
萤火虫的发光细胞含有荧光素和荧光酶， 在该酶的作用下，荧光素与ATP结合 后氧化而发出荧光；求偶信号
The universal energy transporter 每个人每天大约需要消耗45kg ATP, 但每刻存在人体内的
How the enzyme ATP synthase makes ATP？
当H+顺浓度梯度通过 细胞膜时，the F0 part is forced to twist around, 使固定在其上 的F1 也随之旋转发生 构象变化，每一次其中 的一个亚基上催化一 分子ATP的合成。 结合变构机制(binding change mechanism)和旋转催 化假说Boyer （1979）
Paul D. Boyer University of California Los Angeles, USA
John E. Walker Medical Research Council Laboratory of Molecular Biology, Cambridge United Kingdom
F0 c环旋转的荧光显微镜直接观察(1997) Direct observation of rotational catalysis
ATP synthase can work in either direction. If the concentration of ATP is high and the proton gradient low, ATP synthase will run in reverse, hydrolyzing ATP as it pumps protons across the membrane. 该酶还可以反向作用，当 ATP浓度高而H+低时，它 可以水解ATP将H+泵到膜 的另一侧。
活细胞内，可以直接用于
化学反应的能量以化学键形 式储存在三磷酸腺苷
(ATP)中。
ATP广泛存在于
生物细胞中，由
腺嘌呤、核糖 及3个磷酸分子
构成。
热量 生物大分 子的合成 肌肉收缩、细胞 分裂、细胞爬行
胞内或胞 间物质运 输
ATP是一种不稳定的化合物，水解时最外侧的一个高能磷酸
键断裂，同时释放出能量并形成较ATP更为稳定的腺苷二磷酸
Using the H+ gradient to drive the catalytic(催化机制) machinery: the role of the F0 part of ATP F0 synthase A model in which proton diffusion is coupled to the rotation of the c ring of the F0 complex. 在ATP合成酶工作过程中，H+的顺浓度梯度跨膜运动是与ATP 合成酶F0底部c亚基环的逆时针旋转相偶联的。
ATP History ATP first discovered in 1929 by the German chemist Karl Lohmann. Its structure is established some years later. Vladimir Engelhart (Russia) notes in 1935 that muscle contractions require ATP. Between 1939 and 1941 Fritz Lipmann (USA) shows that ATP is the main bearer of chemical energy in the cell . He coins the phrase "energy-rich phosphate bonds". In 1948 Alexander Todd (UK) synthesized ATP chemically.
ATP synthase结构
It consists of an F0 part bound in the membrane and a projecting F1 part. 牛心脏线粒体的电镜 照片：球状颗粒通过 柄附着在线粒体嵴膜 上。(1964)
分离的大鼠肝脏线粒 体ATP合成酶的电镜 照片(1979)
The Nobel Prize in Chemistry 1997
½ Paul D. Boyer and John E. Walker “the enzymatic mechanism underlying the synthesis of adenosine triphosphate (ATP)” ½ Jens C. Skou “the first discovery of an ion-transporting enzyme, Na+/K+-ATPase”
细胞外
细胞内
每水解一个ATP将细胞内的3个Na+泵出细ห้องสมุดไป่ตู้外，同时输入2个 K+维持质膜两侧的离子浓度梯度
Na+/K+-ATPase
Ca2+-ATPase X- represents proteins, which have a net negative charge at the neutral pH of blood and cells