UW Researchers Create Recombinant Viral Vector Repository to Facilitate Biomedical Research

University of Wyoming researchers played a major role in the creation of a key resource designed to facilitate worldwide biomedical research in glycobiology, which focuses on structural and functional analysis of complex carbohydrates.

These molecules are ubiquitous in nature; can function as either free molecules or decorations of larger macromolecules; and impairment of their biological functions can have a major impact on human health.

Don Jarvis, a UW professor in the Department of Molecular Biology, and his colleagues created a repository of recombinant viral vectors encoding 339 different glycoenzymes involved in complex carbohydrate biosynthesis or degradation, which is now available worldwide through the Repository of Glyco-Enzyme Expression Constructs. Viral vectors are tools commonly used by molecular biologists to deliver genetic material into cells.

“The use of these enzymes will lead to discoveries that will reveal underlying causes of certain types of human diseases and ways to treat them,” Jarvis says. “For example, there is a clear correlation between the presence and absence of certain types of complex carbohydrates and certain types of cancer.” 

The new study is highlighted in a paper, titled “Expression System for Structural and Functional Studies of Human Glycosylation Enzymes,” published today (Dec. 18) in Nature Chemical Biology, a monthly, peer-reviewed international journal that publishes research and commentary at the interface of chemistry, biology and allied disciplines.

Kelley Moremen, a professor of biochemistry and molecular biology at the University of Georgia’s Complex Carbohydrate Research Center, and Jarvis were co-lead authors of the paper. Other UW contributors to the paper are Melissa Stuart, a research scientist in molecular biology; Christoph Geisler, a scientist in Jarvis’s startup company, GlycoBac LLC, in Laramie; and Gagan Gahlay, during her previous service as a UW postdoctoral researcher.

“This project was in direct response to a need actively voiced in the glycobiology community,” Jarvis says. “That need was for recombinant enzymes involved in the biosynthesis and degradation of complex carbohydrates, which could be used to study the structure of these enzymes and their products.”

Jarvis and Moremen assembled a large team that eventually created the tools needed to produce the recombinant “glycoenzymes.” The team included investigators from Arizona State University’s Biodesign Institute, which houses a human gene collection that facilitated the project.

These genes were shipped to Jarvis’s lab at UW and Moremen’s lab at the University of Georgia. Jarvis’s group engineered an insect virus to express each gene. These recombinant viruses were then used to infect insect cells, which are routinely cultivated in the Jarvis lab. During the viral infection, the cells produced each of the recombinant glycoenzymes. Moremen’s group used a similar process designed to induce glycoenzyme production in mammalian (human) cells.

Many enzymes were produced at high yields and at similar levels in both hosts, but individual protein levels varied widely. This repository will be a transformative resource for recombinant enzyme production, broadly enabling structure-function studies and expanding applications of these enzymes in glycochemistry and glycobiology, according to the paper.

This latest paper reports the results of basic research designed to establish the glycoenzyme repository, which took place from 2009-2015 with funding from the National Institutes of Health, says Jarvis, who adds this funding and the ability to produce these recombinant glycoenzymes led to the publication of four other papers co-written by his group and several more co-written by the Moremen group.

“Over the course of this seven-year project, we produced about a dozen papers,” Jarvis says.

Other researchers who contributed to the Nature Chemical Biology paper are from Arizona State University’s Biodesign Institute; the Northeast Structural Genomics Consortium at Columbia University; and the Institute for Cell and Molecular Biosciences at Newcastle University in the United Kingdom.