Research

Research projects in the Knight lab span the disciplines of chemistry, biology, and physics, including molecular cloning, bioinformatics, cellular imaging and secretion assays, protein purification, absorbance and fluorescence spectroscopy, stopped-flow kinetics, EPR and NMR spectroscopy, mass-spectrometry based proteomics, Western blotting, and single-molecule optical microscopy. Individual projects use various combinations of these techniques, and many involve collaboration with other groups on our campus and elsewhere. There are opportunities in our group for students with a broad range of interests. Past Knight lab students include majors in Chemistry, Biochemistry, Biology, Psychology, and Engineering. After graduation, our alumni have continued to careers in chemical and biomedical research, medicine, dentistry, veterinary medicine, education, clinical lab science, and science writing. If you are a current or prospective CU Denver student interested in joining the Knight lab, please email Dr. Knight or stop by his office to find out about current opportunities.

Research Topics

Ribbon diagram structure of a portion of the synaptotagmin-7 protein showing two C2 domains connected by a short unstructured linker.

Synaptotagmin-7: ultrasensitive calcium sensor

Synaptotagmin-7 (Syt-7) is among the most Ca 2+ -sensitive synaptotagmins. It is one of two Ca 2+ -dependent synaptotagmins (along with Syt-9) that are abundant in pancreatic β-cells. Ca 2+ -dependent synaptotagmins bind Ca 2+ and membranes through their C2A and C2B domains, located at their C-terminus. In 2018, our...
Diagram showing how synaptotagmin-like proteins help insulin secretory vesicles dock to the β-cell plasma membrane via their two C2 domains.

Synaptotagmin-like proteins: calcium-independent C2 domains that target anionic membranes

Update 11/30/2023: Congratulations to Nara on her paper being accepted into Protein Science! You can access the pre-print either on the Protein Science website or at BioRxIV . The synaptotagmin-like protein (Slp) family comprises five mammalian proteins (Slp-1 through Slp-5) that function as Rab27 effectors to tether secretory vesicles to...
Graph of mass spectra showing a peak at 17645 Daltons for unmodified protein and 17903 Daltons for modified protein.

Post-translational modification of C2 domains: role in oxidative stress?

Biochemistry students learn that proteins are composed of the 20 biological amino acids linked together by peptide bonds. In reality, many of these 20 amino acids can also be chemically modified inside of cells, creating a profound diversity of possible structures. In addition to the many known enzyme-catalyzed post-translational modifications...
The lateral diffusion constants are 2.7 square microns per second for the monomer, 1.4 for the dimer, and 0.9 for the trimer.

Single-molecule diffusion: a powerful tool for investigating the stoichiometry of protein-lipid interactions

One of the major scientific advances of the late 20th and early 21st century was the ability to see individual protein molecules in real time using single-molecule fluorescence microscopy. In his postdoctoral research from 2006-2010, Dr. Knight pioneered the use of single-molecule fluorescence microscopy to observe individual fluorescent-tagged proteins binding...
Stubs are made by adsorbing and rupturing liposomes onto a planar glass surface, washing with water, and adding physiological buffer.

Supported Tubulated Bilayers (STuBs): a new method for preparing lipid tubules

Most membranes in a cell are highly curved, not flat. Yet, most of the experimental methods for studying lipid membranes are either flat (e.g., supported lipid bilayers) or have very low curvature (e.g., large unilamellar vesicles). In order to support the study of proteins binding and acting on curved membranes,...
Diagram of a student-centered literature review writing assignment and class presentation about a cell signaling or metabolic pathway.

Advancing Biochemistry and Biophysics Education

In an effort to constantly improve university science education, Dr. Knight has collaborated with students and faculty to publish articles in the education literature. From 2014-2018, we teamed up with Honors General Chemistry (HGC) professor Dr. Peggy Bruehl and library resource specialist Denise Pan to study how teaching information literacy...