Protein Crystallization. Techniques, Strategies, and Tips Edited by Terese M. Bergfors. International University Line, 1999. ISBN 0963681753.
Protein Crystallization. Techniques, Strategies, and Tips Edited by Terese M. Bergfors. International University Line, 1999. ISBN 0963681753.
复制标题
蛋白质结晶。
DOI：
10.1021/cg020059c
复制
发表时间：
2003
影响因子：
3.8
通讯作者：
J. Glusker
中科院分区：
文献类型：
--
作者：
J. Glusker
这是一本需要躺在每个实验室的工作台上的书，在那里试图生长蛋白质晶体。在高通量晶体学的今天，它的出版时机是完美的。它基本上包含了一套生长晶体的实验方案，如果你的第一次尝试没有产生晶体，应该改变什么的提示，以及为什么使用每种方法的一些解释。目前使用的所有方法都是用简单的术语描述的，在许多情况下，在实验室中进行的实验很简单。读者预计不会有太多的经验，在增长晶体，所以这是一个很好的书使用，如果你正在开始增长晶体。然而，它也提供了其他策略的想法，如果你的结晶实验没有工作，可以尝试。编辑是来自瑞典乌普萨拉大学的Terese Bergfors，他有相当丰富的教学经验。她做了一项了不起的工作，首先确定了蛋白质晶体学家需要的一本书，其次以这样一种方式编辑它，即她保持了每章相似的风格和内容，即使她只写了一些章节。这是任何编辑多作者卷的人的模型;它读起来无缝，就好像只有一个作者。共19章，16位作者。编辑已经选择了几种新的技术以及旧的尝试和真实的（她在许多情况下提供了替代方法）。提供了大量的信息，分为简短的章节，是准确和中肯的。实验技术被很好地描述，并且在每一章的末尾有许多相关的参考文献，以便可以很容易地找到额外的信息，并且谢天谢地，文章的标题也被列出，这使得读者更容易选择所需的。这本书以亚历克斯·麦克弗森（Alex McPherson）的《关于结晶蛋白质的一点建议》（A Bit of Advice on Crystallizing Proteins）一章开始。他指出，总的目标是说服生物分子，“它们在晶体中比在溶液中更好，如果你愿意的话，更快乐。”麦克弗森描述了结晶的两个阶段。首先，"...分子必须克服能量障碍，形成临界尺寸的周期性有序聚集体，这种聚集体在热力学意义上可以“生存”。第二，如果人们能够使“固态比自由的溶液态对单个分子更有吸引力”，就可以实现增长。然后概述了实现这些目标的方法。然而，在书的最后，亚历克斯卡梅隆指出了这一点（在题为“最后一条建议：外表可能是骗人的！”的一章中）最美丽的晶体不一定是最好的衍射器，当然，它是一个良好的衍射图案是最终目标。最初这本书涵盖了基本技术，材料和参数涉及成功的结晶。建议使用动态光散射法测定蛋白质的粒度分布和聚集程度，从而测定其结晶性。强调蛋白质纯度。“自己纯化蛋白质总是有利的，因为你在这个过程中了解了很多关于它的行为。然而，如果你自己没有净化它，就会给出建议。实用的化学技术，如使用沉淀剂（列表），找到最佳的pH值和结晶温度，结晶筛的类型（稀疏矩阵和其他），条纹播种，和macroseeding包括在内。结晶策略在几个章节中涉及，即如何计划工作。有章节的结晶cryoctylallography，生长的晶体下石油，以及如何做.
This is a book that needs to lie on the benches of every laboratory in which attempts are being made to grow protein crystals. In these days of high-throughput crystallography, the timing of its publication is perfect. It essentially contains sets of experimental protocols for growing crystals, hints on what to change if your first try does not yield crystals, and some explanations of why one uses each method that has been written about. All of the methods that are currently used are described in simple terms and, in many cases, with experiments that are simple to follow in the laboratory. The reader is not expected to have had much experience in growing crystals, so this is a good book to use if you are starting to grow crystals. However, it also provides ideas on other strategies that can be tried if your crystallization experiments have not worked. The editor is Terese Bergfors from Uppsala University, Sweden, who has considerable experience teaching the subject. She has done a marvelous job, first by identifying a book that protein crystallographers need, and second by editing it in such a way that she has maintained a similar style and contents for each chapter, even though she only wrote some of the chapters. This is a model for anyone who edits a multiauthor volume; it reads seamlessly as if there was only one author. There are 19 chapters covered by 16 authors. The editor has selected several new techniques as well as the old tried and true ones (for which she offers alternative methods in many cases). Much information is provided, divided up into short chapters that are precise and to the point. Experimental techniques are well-described, and there are many relevant references at the end of each chapter so that additional information can readily be found, and thankfully, the titles of the articles are also listed, which makes it easier for the reader to select the required ones. The book begins with a chapter entitled “A Bit of Advice on Crystallizing Proteins” by Alex McPherson. He points out that the overall aim is to persuade biological molecules that “they are better off, happier if you will, in the crystal than in solution.” McPherson describes the two phases of crystallization. First,“... molecules must overcome an energy barrier to form a periodically ordered aggregate of critical size that can “survive” in a thermodynamic sense.” Second, growth is achieved if one can make “the solid state more attractive to individual molecules than the free, solution state.” Methods for making these happen are then outlined. In the end of the book, however, it is pointed out by Alex Cameron (in a chapter entitled “One last piece of advice: appearances can be deceiving!”) that the most beautiful crystals are not necessarily the best diffractors and, of course, it is a good diffraction pattern that is the ultimate aim.Initially the book covers basic techniques, materials, and parameters involved in successful crystallization. Dynamic light scattering is recommended for determination of the size distribution and degree of aggregation of the protein, hence its crystallizability. Protein purity is stressed.“It is always advantageous to purify the protein yourself because you learn a great deal about its behavior in the process.” However, advice is given if you did not purify it yourself. Practical chemical techniques such as which precipitants to use (with lists), finding the optimal pH and temperature for crystallization, types of crystallization screens (sparse matrix and others), streak seeding, and macroseeding are included. Crystallization strategy is covered in several chapters, that is, how to plan the work. There are chapters on crystallization for cryocrystallography, the growth of crystals under oil, and what to do …