Engineering topics

Miller, Jacob

Publications and source records attributed to Miller, Jacob.

2 records

DOE OSTI2023

Cancer-associated somatic mutations in human phosphofructokinase-1 reveal a critical electrostatic interaction for allosteric regulation of enzyme activity

Metabolic reprogramming, including increased glucose uptake and lactic acid excretion, is a hallmark of cancer. The glycolytic ‘gatekeeper’ enzyme phosphofructokinase-1 (PFK1), which catalyzes the step committing glucose to breakdown, is dysregulated in cancers. While altered PFK1 activity and expression in tumors have been demonstrated, little is known about the effects of cancer-associated somatic mutations. Somatic mutations in PFK1 inform our understanding of allosteric regulation by identifying key amino acid residues involved in the regulation of enzyme activity. Here, we characterized mutations disrupting an evolutionarily conserved salt bridge between aspartic acid and arginine in human platelet (PFKP) and liver (PFKL) isoforms. Using purified recombinant proteins, we showed that disruption of the Asp–Arg pair in two PFK1 isoforms decreased enzyme activity and altered allosteric regulation. We determined the crystal structure of PFK1 to 3.6 Å resolution and used molecular dynamic simulations to understand molecular mechanisms of altered allosteric regulation. We showed that PFKP-D564N had a decreased total system energy and changes in the electrostatic surface potential of the effector site. Cells expressing PFKP-D564N demonstrated a decreased rate of glycolysis, while their ability to induce glycolytic flux under conditions of low cellular energy was enhanced compared with cells expressing wild-type PFKP. Taken together, these results suggest that mutations in Arg–Asp pair at the interface of the catalytic-regulatory domains stabilizes the t-state and presents novel mechanistic insight for therapeutic development in cancer.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

United States cool surfaces deployment plan

Solar-reflective building envelope surfaces, such as cool roofs and walls, can be especially helpful in disadvantaged communities that often have poorly insulated older homes, aging or absent air conditioning units, steep utility bills, polluted air, and high vulnerability and exposure to extreme heat. With support from the U.S. Department of Energy, our project seeks to dramatically increase the climate-appropriate deployment of cool surfaces across the United States with an emphasis on their application to disadvantaged communities. First, we sought to identify cool-surface deployment barriers, opportunities, and models by (a) reviewing the history of cool-surface deployment activities, (b) interviewing cool-surface stakeholders, (c) researching successful energy-efficiency/green building deployment models; and (d) interviewing the actors who have implemented these models. Second, we conducted a workshop to engage stakeholders in development of a deployment plan. Third, we asked several U.S. federal agencies (a) how cool surfaces and cool surface stakeholders could support their missions and (b) how agency activities could support cool-surface deployment. Fourth, we identified a set of transformative ideas that form the core of the deployment plan. Transformative ideas include but are not limited to initiatives to (a) launch an educational campaign to make the general public and building professionals aware of how cool roofs and walls exclude unwanted solar heat; (b) create a “Cool Roof Prize” stimulating the development of affordable, high-performance cool asphalt roofing shingles; (c) conduct high-profile, large-scale demonstration programs that bring cool surfaces to disadvantaged heat-vulnerable communities; and (d) support local, regional, and state climate action (heat mitigation) plans with cool surfaces.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗