Targeted Protein Degradation: A Bright Future for Drug Discovery

targeted protein degradation and protacs

Our cells have evolved multiple mechanisms for “taking out the trash”—breaking down and disposing of cellular components that are defective, damaged or no longer required. Within a cell, these processes are balanced by the synthesis of new components, so that DNA, RNA and proteins are constantly undergoing turnover.

Proteins are degraded by two major components of the cellular machinery. The discovery of the lysosome in the mid-1950s provided considerable insight into the first of these degradation mechanisms for extracellular and cytosolic proteins. Over the next several decades, details of a second protein degradation mechanism emerged: the ubiquitin-proteasome system (UPS). Ubiquitin is a small, highly conserved polypeptide that is used to selectively tag proteins for degradation within the cell. Multiple ubiquitin tags are generally attached to a single targeted protein. This ill-fated, ubiquitinated protein is then recognized by the proteasome, a large protein complex with proteolytic activity. Ubiquitination is a multistep process, involving several specialized enzymes. The final step in the process is mediated by a family of ubiquitin ligases, known as E3.

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New Year, New Buildings, Same Commitment to Sustainability

In times of rapid growth, we look to the future with excitement while also assuring that our expansion is sustainable. The Promega Global Facilities Planning Team emphasizes environmental stewardship and long-term planning. Each building is designed to meet ambitious sustainability goals, and innovations incorporated into each project inform the next. In 2019, we finished construction on two new buildings in Europe and made progress on two important facilities at our headquarters in Wisconsin.

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Sustainability Makeover: Parking Ramp Edition

parking_ramp_solar_panels

As Promega grows globally and locally here at headquarters, the construction of new and expanding facilities are considered with great care to ensure a commitment to sustainability. Between April and November of 2019, the parking ramp located near the Feynman Center received a massive upgrade with long-term impacts on the company’s sustainability goals.

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Enhancing Creativity: The Promega Employee Art Show 2020

What hangs in the hallways of your workplace? Advertising? Awards? Commercially-sourced artwork? The Promega campus in Madison, WI, is composed of eight buildings. In many of these buildings you’ll find all of the above.

But as you enter the atrium of the BTC (Biopharmaceutical Technology Center) building where the Employee Art Show hangs, you’re greeted by handmade, homemade artwork, including drawings, ceramics, paintings, photographs and quilts.

Promega employees examine artwork in the BTC during the 2020 Employee Art Show opening.

These art pieces hold the distinction of being created by talented Promega employees, their families and friends. The annual Promega Employee Art Show opened Friday, January 17, with approximately 150 works displayed, including pieces created by parents of employees, employees and their children. These art pieces are on display to the public through the end of February 2020.

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Stakeholder Capitalism—40 Years Later

Today’s blog is written by guest blogger, Penny Patterson, VP Corporate Communications at Promega.

The idea that businesses need to serve and provide value to constituents in addition to shareholders is one that has gained increasing recognition since last summer when the Business Roundtable issued its “Statement on the Purpose of the Corporation.”   The topic of what some call “stakeholder capitalism” is surfacing again heading into the World Economic Forum this week.

Promega has practiced “stakeholder capitalism” for more than 40 years and, as we’ve shared through our corporate responsibility reporting for the last decade, we have seen meaningful impact. From our founding in 1978, we have taken a “whole human” approach to our business. For us this means growing a financially stable and profitable company that considers and benefits science, employees, customers, community, shareholders and all global residents.

This approach starts with our people. We live the notion that every one of our employees has the potential to make a meaningful difference. And they do. Here are just a few examples. Our manufacturing and operations teams deliver with 99% accuracy and a complaint rate of 0.004%.  Discoveries by our R&D scientists generate some of the most read papers among key science journals. The average tenure of our leadership team is 18 years, and over half of these leaders grew their careers and capabilities at Promega.

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Popular Papers from Promega Authors

Promega is a chemistry and instrument supplier to scientists in diverse industries and research labs around the world. True. But we are more than just a supply company; we are scientists dedicated to supporting the work of other scientists. We want the science behind the technologies we develop to be both vetted and valued by the scientific community at large, which is one reason our scientists take the time to prepare and submit manuscripts to peer-reviewed journals. Here we call out some of our published research papers that were highly read in 2019. In the journal ACS Chemical Biology alone, five Promega-authored papers were among the top 10 most read papers in 2019. Here’s a quick review of the highlights from these ACS papers.

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Green Chemistry is Better Chemistry

When you think of sustainability, what comes to mind? Immediately, my brain imagines vast collections of plastic in the ocean and carbon emissions from millions of cars. I’m guessing that, like me, you didn’t think about optimizing the synthesis of chemical reactions to reduce toxicity or energy usage. Although we’re often focused on the more visible forms of waste, sustainability applies to an enormous range of human activities.

Promega is committed to integrating the principles of sustainability across all aspects of our business. One recent area of focus for our PBI branch is a shift toward Green Chemistry. PBI synthesizes reagents and small molecules used in Promega products. After deciding “it was the right thing to do for our customers and for the environment,” the leader of Promega’s Corporate Responsibility Program, Corey Meek, assembled a few individuals to start a conversation about implementing Green Chemistry principles.

“It was the right thing to do for our customers and for the environment.”

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Learnings From the Eppendorf Exchange Program

Last year, on Promega’s 40th anniversary, we received a generous gift from a friend in the industry: Eppendorf. That gift was an exchange program. The teenage child of any Promega employee was given the opportunity to visit an Eppendorf family in another country, and in return host the Eppendorf family’s child in their home. The goal was for both children to experience another culture and build a relationship with each other.

In 2019, 11 Promega children bid good-bye to their parents, hopped on a plane, and flew to Germany. There they would stay for three weeks with a family they’ve never met. For all involved, it proved to be a valuable and positive learning opportunity. Here are a few takeaways from their experience:

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Researching the Researchers: Aberto Biscontin

2019 Real-Time PCR Grant

The three 2019 Real-Time PCR Grant Winners have been hard at work in the six months since winning their grants. Each winner was eligible to receive up to $10,000 in free PCR reagents as well as the opportunity to collaborate with our knowledgeable technical service and training teams.

One of the 2019 winners, Alberto Biscontin (University of Padova, Italy), performs research in the fields of Neurogenetics and Chronobiology. He is looking to shed greater light on the circadian rhythms of the Antarctic krill. Alberto published his most recent analysis in Nature and GoTaq® qPCR Master Mix helped him validate expression of genes for his study.

His qPCR data showed support for internal mechanisms that not only support daily living but also clarified the overwintering process of the krill. Now that Alberto has sized up some zooplankton, we asked him to share a little more about himself and his research:

Q: How long have you been a researcher?
A: I have been a researcher since 2012.

Q: How did you decide to research Antarctic krill?
A: In 2013, I had the opportunity to join the international Antarctic research program PolarTime. [It] brought together eight research groups with different scientific expertise to study seasonal and daily rhythms in the Antarctic krill Euphausia superba.

Q: When you are not busy at the bench, what do you like to do?
A: Traveling. I love strolling through open-air markets.

Q: Are there any tips or tricks you have learned that make your job easier?
A: You can easily switch from a classic RT-PCR protocol to a cheaper and faster One-step protocol using the same primers and temperatures.

Q: What comes next?
A: I would like to characterize the clock machinery of other polar organisms to understand whether high latitude clocks have developed similar strategies to cope with [the] polar environment. Moreover, a better understanding of marine circadian clocks could help to shed light on the evolution of the animal circadian machinery.

You can find Alberto’s most recent publication in Nature Scientific Reports. The 2020 Real-Time PCR Grant will be coming soon. For more information on the 2019 winners and information on the 2020 Grant, visit the Real-Time Grant web page. Be sure to follow us on social media for the most up-to-date information regarding the 2020 Grant, including application deadlines and winner notifications!

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Cloning with pGEM®-T Vectors: Ligation

One of the easiest methods for cloning blunt-ended DNA fragments including PCR products is T-vector cloning, such as with pGEM®-T or pGEM®-T Easy Vector Systems. This method takes advantage of the “A” overhang added by a PCR enzyme like Taq DNA Polymerase. T vectors are linearized plasmids that have been treated to add 3′ T overhangs to match the A overhangs of the insert. The insert is directly ligated to the T-tailed plasmid vector with T4 DNA ligase. The insert can then be easily transferred from the T vector to other plasmids using the restriction sites present in the multiple cloning region of the T vector.

Proofreading polymerases like Pfu do not add “A” overhangs so PCR products generated with these polymerases are blunt-ended. In a previous blog, we discussed a simple method for adding an A-tail to any blunt-ended DNA fragment to enable T-vector cloning. Below, we think about the next step: Ligation.

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