What do structural biologists do?

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  • Опубліковано 23 гру 2024

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  • @thebumblingbiochemist
    @thebumblingbiochemist  Рік тому +5

    *mutation technically refers to changes at the gene level, but we commonly refer to the resultant changes in the protein as mutations as well. But some people might get irritated if you do so be forewarned!
    Although you can’t “mutate” proteins inside a person, you can give a person mutated proteins! Don’t worry - mutations can be good! Based on structures of insulin, for example, scientists have been able to make modified versions that are longer-lasting or faster-acting. Much more on how here: blog form: bit.ly/insulin_diabetes_biochem ; UA-cam: ua-cam.com/video/Mp8jUqfjMBs/v-deo.html
    Those are a couple examples of “translational” research, what we call research that has a direct applicational benefit. But structural biology is also crucial to “basic” research, where we focus primarily on trying to figure out how things work (and this can lead to important applications, etc. down the line).
    Structures can guide crucial experiments and what’s especially awesome about them is that they are freely accessible through the PDB (Protein DataBank) for anyone to use to inform their own experiments - or just to peruse!
    If you want to learn more about how: intro to PDB, crystal structure entries, crystal contents, resolution: bit.ly/pdbstructures & ua-cam.com/video/Re2gwi-_OEw/v-deo.html & ua-cam.com/video/IZtHsUFbyes/v-deo.html      
    But before I make this all seem too simple… getting a structure usually involves a LOT of hard work. Sometimes by multiple people. Because (assuming you are trying to solve the structure of a protein or proteins that you express recombinantly, not ones you purify in their “native” state from cells) you have to…
    - clone the protein(s) (stick their genetic recipes into a form you can work with, usually a circular piece of DNA that you can stick into cells)
    - Express LOTS of the protein(s) (stick that cloned DNA into cells like bacteria, insect, yeast, or mammalian cells and get them to make it)
    - [Figure out why the cells don’t want to express your protein… maybe change the construct (version of the recipe)… use a different cell type…]
    - Purify the protein(s) SUPER well (typically using multiple column chromatography steps)
    - [Figure out why it isn’t getting very pure despite your attempts at it…]
    - Prepare complexes if applicable (mix proteins together, etc.)
    - Prepare the samples for data collection (vitrify grids for cryo or crystallize the protein for crystallography)
    - [Do TONS of screening of different conditions to try to get the protein to crystallize… go back to the start and try altering the construct, maybe removing regions predicted to be disordered]
    - For cryo-EM you often start by screening things with a low-resolution technique like negative staining or a screening microscope, then optimize conditions before going for the big time collection
    - Collect LOTS of HIGH QUALITY data
    - [start over to get better data…]
    - Process all that data
    - Generate a model based on that data
    - Refine the model
    - [these last couple parts involve a lot of time at a computer]
    And that’s “just” to get the structure! Then comes the analysis, functional follow-up studies etc.
    Note: the workflow will differ depending on the project and the experimental systems you use - more on experimental systems here: bit.ly/experimentalsystems & ua-cam.com/video/fewMfNw9Je0/v-deo.html
    Hope that helped give you a flavor of structural biology - I know it can seem really obscure, but it’s super useful. And super cool. And involves more than “just” solving structures (which IS a super important - and hard - part of it).
    To review:
    * The what: Structural biology is a scientific discipline that looks at the molecular structure of biological macromolecules and how that STRUCTURE relates to its FUNCTION
    * The why: To answer questions like:
    * Why do molecules work the way they do?
    * What specifically makes one (or a group of them) well-suited for a particular task?
    * Can we manipulate them to work even better or do other things?
    * The how:
    * incorporates principles and techniques of: biochemistry, biophysics, molecular biology
    Bottom line: Structure & function are intimately connected. We can exploit this relationship to learn about function from structure and structure from function. It’s hard, but fun and often rewarding, work.
    For more structural-biology content, I created a page on my blog with links to my past posts on it: bit.ly/structural_biology  

    and here's a link to my UA-cam structural biology playlist: ua-cam.com/play/PLUWsCDtjESrGhwVxsRbTJdL-BEsN60RCs.html  

    and here's a link to my UA-cam structural biology playlist: ua-cam.com/play/PLUWsCDtjESrGhwVxsRbTJdL-BEsN60RCs.html  

    Also, for all that protein expression and purification: 
    - blog page: bit.ly/proteinpurificationtech
    - UA-cam playlists 
    - recombinant protein expression & purification: ua-cam.com/play/PLUWsCDtjESrHrxSeKsNxMlooYe0NwEQbV.html
    - protein chromatography: ua-cam.com/play/PLUWsCDtjESrEtqQkEXsQpTyRfMhywBVQ7.html
    more on site-directed mutagenesis: bit.ly/mutagenesisconstructs & ua-cam.com/video/vm6p5Nq2sTg/v-deo.html
    more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 bit.ly/2OllAB0 or search blog: thebumblingbiochemist.com

  • @sohinisaha4069
    @sohinisaha4069 10 місяців тому

    I have recently gotten interested in structural biology and various techniques used in studying proteins. Your channel is wonderful. I love every video you post

    • @thebumblingbiochemist
      @thebumblingbiochemist  10 місяців тому

      Thank you so much! Best of luck! There's definitely a bit of a steep learning curve for structural biology but it's a lot of fun once it clicks!

  • @oromhal
    @oromhal 11 місяців тому +1

    Thank you so so much! Such a helpful video for understanding what is Structural Biology!

    • @thebumblingbiochemist
      @thebumblingbiochemist  11 місяців тому +1

      I'm so glad it was helpful! I never knew before grad school what it was!

  • @sabrinakurtze7438
    @sabrinakurtze7438 2 місяці тому

    Hi! This is a great video! I am very interested in going into the field of structural biology but am not sure what major to go into for that. What major would you suggest I study for my bachelor's since I would like to learn structural biology?