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Single chip tests thousands of enzyme mutations at once - Nature.com

Single chip tests thousands of enzyme mutations at once - Nature.com

Single chip tests thousands of enzyme mutations at once - Nature.com
Jul 23, 2021 2 mins, 13 secs

Figuring out how a protein or enzyme works, and understanding how genetic mutations affect these molecules that are fundamental to life, can often take years.

Researchers must alter hundreds of the molecule’s amino acid building blocks one-by-one, produce each mutated enzyme in the lab and test how each mutation affects the enzyme’s ability to carry out its job.

Now, a glass chip etched with tiny channels could reduce that time to mere hours by allowing researchers to test more than a thousand mutations at a time.

To develop HT-MEK, bioengineer Polly Fordyce and biochemist Daniel Herschlag at Stanford University in California and their colleagues worked for six years, ending up with a US$10 chip about 7 cm2 in size.

To test the system, Fordyce and Herschlag chose a bacterial enzyme called PafA that is involved in modifying other proteins.

A scanner measured the amount of light given off by the chemical: mutations that made PafA less effective made the enzyme produce less light.

Rather than simply telling the researchers whether the experiment worked or not, the platform allowed them to examine the speed at which each mutant enzyme carried out the reaction and determine how chemicals or pH changes affected the way the enzyme folds and functions.

Because it can screen so many mutants at a time, the system could allow researchers to look beyond mutations in the active site — the part of an enzyme that actually carries out its main function and usually attracts the most research attention.

The extent of the mutations’ impact was surprising, says Herschlag, who has spent many years studying the enzyme.

He and Fordyce say that being able to identify the functions of these distant mutations might allow researchers to target enzymes that are considered ‘undruggable’ because their active sites are structurally similar to those of other, healthy enzymes.

“It’ll be exciting to see where this technology goes, the scale-up here is pretty impressive.” He expects that HT-MEK will make many tasks easier and faster, but that it remains to be seen whether the system will work as well for every type of enzyme as it did for PafA.

Although the instructions for building a HT-MEK system have been published online2, Fordyce and Herschlag hope to create a centre where researchers can come to test the enzymes they’re interested inJ

Herschlag says he is especially excited about the potential to understand how certain genetic mutations lead to diseases and how these mutant enzymes can be targeted with drugs.

If researchers can analyse the functions of these mutations more quickly, he says, “we'll be creating the knowledge necessary for going from the molecular changes to ultimately predicting disease outcomes”.

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