Journal club: structure of the ternary Prp31:15.5K:U4 snRNA complex (1 reply)
I would like to start the “journal club” section of the NMR forum by presenting my favorite paper from the recent literature.
The paper (in this weeks issue of Science) is, in my opinion at least, a perfect example of the ability of NMR to contribute to the structural analysis of a challenging RNA-protein
complex
I chose this high-impact paper because the combination of biochemistry, NMR, and crystallography on a ternary complex is how I would like to tackle a challenging project that I am working on.
The authors used chemical shift perturbation and saturation transfer to monitor the effect of titrating Prp31 into the preformed U4-15.5K RNA-protein complex. To do this the authors purified nitrogen-15 labeled and deuterated samples of the 15.5K construct. All of the proteins were produced in E.coli using well-established protocols. Gel filtration was used to purify the complexes to yield homogeneous samples
Subsequent crystallogrpahic analysis confirmed formation of the ternary complex and provides a rationale for how Prp31 contributes to the stability of the complex. The structural studies in combination with biochemical experiments yield valuable insights into spliceosome mechanism. NMR experiments such as the ones reported here are well within the reach of most laboratories with access to instrumentation and can be very powerful in lending confidence to subsequent crystallization experiments.
Your comments are welcome!
Michael.
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