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I’m always on the lookout for new ways of teaching proteomics. Here’s a gem that I found on YouTube.
Directed in 1971 by Robert Alan Weiss for the Department of Chemistry of Stanford University and imprinted with the “free love” aura of the period, this short film continues to be shown in biology class today. It has since spawn a series of similar funny attempts at vulgarizing protein synthesis. Narrated by Paul Berg, 1980 Nobel prize for Chemistry.
Protein complexes in a muscle cell. Image provided by RUB. © Prof. Wolfgang A. Linke
While we are all familiar with the role of methyltransferase in DNA and protein modification in the nucleus, (think epigenetics with regards to DNA), this is the first time that methylation in the cytoplasm has been shown to promote protein complex formation.
The researchers first identified an enzyme which is mainly present in the cytoplasm and which methylates the amino acid lysine (Smyd2). Then they searched for interaction partners of the enzyme Smyd2
and found the heat shock protein Hsp90. The scientists went on to show that Smyd2 and methylated Hsp90 form a complex with the muscle protein titin.
According to the authors, “Titin is the largest protein in the human body and known primarily for its role as an elastic spring in muscle cells. Precisely this elastic region of titin is protected by the association with methylated Hsp90.”
In skeletal muscle cells of the zebrafish, the team explored what happens when the protection by the methylated heat shock protein is repressed. By genetic manipulation they altered the organism in such a way that it no longer produced the enzyme Smyd2, which blocked the methylation of Hsp90. Without methylated Hsp90, the elastic titin region was unstable and muscle function strongly impaired; the regular muscle structure was partially disrupted.
Click here for a link to the Genes and Development paper.
So you’ve isolated your protein, ran them on a gel and now you’re ready to transfer them to a membrane to begin western blotting. Sounds simple, right? Not so fast. Don’t forget to equilibrate your gel prior to beginning your transfer. Gel equilibration generally involves rinsing the gel in diH2O and soaking it in transfer buffer for approximately 15 min. While it may sound simple, (and it truly is), it is a step that might make the difference between an ugly blot and one that is publication worthy.
Below are some points to consider about gel equilibration:
To learn more tips and tricks, download the Protein Blotting Guide from Bio-Rad Laboartories.
Despite the difficulties associated with measuring in-situ protein-protein interactions in neural networks, Dr. Akira Chiba of the University of Miami recently announced that his team has embarked on a project to develop a protein interaction map within brain cells. Why have these studies been so difficult to perform until now and what does Dr. Chiba have that will make him successful? The answer lies in the small size of neural proteins and the technical limitations associated with even the highest resolution microscope.
Using a a custom- built 3D FLIM (fluorescent lifetime imaging microscopy), Chiba’s team has been able to spatially and temporally quantify fluorescently tagged protein-protein interactions in genetically modified fruit flies.
According to Chiba, “collaborating fluorescent chemistry, laser optics and artificial intelligence, my team is working in the ‘jungle’ of the molecules of life within the living cells. This is a new kind of ecology played out at the scale of nanometers—creating a sense of deja vu 80 years after the birth of modern ecology.”
Thanks to the University of Miami for this story.
Watch the video below, (already set to the appropriate frame…just press play), as Karl Svoboda of the Howard Hughes Medical Institute explains how FLIM works.
Now that we’ve finished our series on Proteomics Application Tips, it’s time to reward ourselves with a captivating talk by Professor Danny Hillis on Understanding Cancer Through Proteomics. Yes…he’s preaching to the choir, but it’s still fun to see our cause promoted on the “big screen.” Enjoy!
