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X射线晶体学研究热木体马里蒂玛M42氨基肽TmPep1050的寡聚态过渡
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
DOI:

11:27 min

May 13, 2020

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Chapters

  • 00:04Introduction
  • 01:03Activity Assay
  • 02:19Apoenzyme Preparation
  • 03:32Dimer Preparation
  • 04:03Microseeding
  • 05:01X-Ray Diffraction
  • 05:48Indexation
  • 06:50Molecular Replacement
  • 08:04Model Building
  • 09:07Results: Representative Thermotoga Maritima M42 Aminopeptidase TmPep1050 Oligomeric State Transition
  • 10:50Conclusion

Summary

Automatic Translation

该协议已经开发,以研究TmPep1050的二丁二代过渡,一种M42氨基肽,在结构层面。从蛋白质纯化到 X 射线数据处理,这是一条简单的管道。通过一个案例研究,TmPep1050H60A H307A 变种强调了晶体生成、数据集索引和分子 替代。

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