Studying Mitochondrial Fission with NanoBiT Complementation Assay

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Motivation
It’s a new year. Whether you’re a self-improvement fanatic or just ready for good things to start happening, you’ve got a plan. You might be changing up an old exercise routine or trying a new cooking technique.

And at work, you are digging deeper; this is the year you illuminate the protein interactions that you’ve previously not been able to visualize.

Good news. There is a new protein complementation assay that can help. Studying-mitochondrial-fission-poster

About NanoBiT
NanoBiT™ Complementation Reporter is a recently developed protein interaction assay that features the improved NanoLuc® luciferase. NanoLuc, originally isolated from a deep sea shrimp, is a small luciferase that provides a much brighter signal than firefly luciferase.

About Split Luciferase Systems
If you’re interrogating two proteins to understand the conditions under which they interact, a split luciferase system enables you to tag each protein with a luciferase subunit. Interaction of the tagged proteins facilitates the complementation of the subunits, resulting in a luminescent signal.

However, not all split firefly luciferase systems are created equal. Using NanoLuc luciferase, two NanoBiT subunits have been developed and independently optimized for stability and minimal self-affinity, as well as small size (18kDa for LgBiT and 11 amino acids for SmBiT). The benefits of NanoBiT include:

• small subunits that are not likely to interfere with protein interactions
• a bright signal that results in a greater signal:background ratio and thus a more sensitive assay

A Picture is Worth a Thousand Words
You can see data demonstrating the sensitivity and brightness of NanoBiT here, including data showing NanoBiT detection of protein association and dissociation. You will also find several short chalk talks presented by researchers that are using NanoBiT in protein interaction studies.

One of the chalk talks features Dr. Stefan Strack, an investigator at the University of Iowa Carver College of Medicine, who discusses his studies on mitochondrial fission using a split luciferase. The goal of his work is to learn more about the mechanism of mitochondrial fission.

Mitochondria and Disease
Mitochondria are organelles found in abundance in most cells of plants and animals. Often called the cells’ powerhouse, mitochondria are where cellular respiration takes place; energy in the form of ATP is created when mitochondria intake and metabolize breakdown products of carbohydrates and fatty acids.

Not surprising given their importance in energy metabolism, dysfunction of mitochondria has been associated with serious diseases such as muscular dystrophy, Parkinson’s, Huntington’s and Alzheimer’s disease, as well as conditions that can result in blindness.

Strack’s group is interested in interactions of mitochondrial outer membrane proteins Drp-1 and mff (mitochondria fission factor).

Why Use NanoBiT?
The Strack laboratory decided to try the new NanoBiT™ complementation assay, and realized that the better NanoBiT signal to background ratio compared to the split firefly luciferase, gave them a much more sensitive assay.
In addition, the small size of the NanoBiT subunits means that the Strack team doesn’t have to worry about the two subunit tags interfering with normal mitochondrial protein function.

Watch the Video
Here’s Dr. Strack to discuss his research:

You can learn more about the NanoBiT Complementation Assay, see other videos, view data, and even join a forum to discuss your questions and results with others, including product experts, here.

Join the Conversation
We are always happy to hear from you, and there is a forum on the NanoBiT page that you can join. The forum lets you see what others are doing with NanoBiT; you can add your story there. Product experts are available to answer your questions too.

Let’s make 2016 the year we get things done, together!

Here are the links again:

NanoBiT page

Studying Mitochondrial Fission with NanoBiT Chalk Talk

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Kari Kenefick

Kari has been a science writer/editor for Promega since 1996. Prior to that she enjoyed working in veterinary microbiology/immunology, and has an M.S. in Bacteriology, U of WI-Madison. Favorite topics include infectious disease, inflammation, aging, exercise, nutrition and personality traits. When not writing, she enjoys training her dogs in agility and obedience. About the practice of writing, as we say for cell-based assays, "add-mix-measure".

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