Revealing the Regulatory Role of Mn2+ in diverse biological processes via development and implementation of a biochemical Mn2+ sensor 

Objectives: Characterize the molecular mechanisms of Mn2+ homeostasis. Elucidate the Mn2+ metalloproteome. Create a microfluidic calorimetric Mn2+ sensor.

Team Coordinator: Steven Damo, PhD (Subproject PI)

Collaborators: Nelms (Fisk, biology); N. Arnett (Fisk, chemistry), R. Mu (Fisk, physics), L Qian (Fisk CS); Vanderbilt Collaborators: Walter Chazin (Structural Biology), Eric Skaar (Microbiology/Immunology), and DeYu Li (Biomedical Engineering)

Scientific Impact: This transformative approach leverages the basic biochemistry of the protein calprotectin at the center of an interdisciplinary effort bridging structural biology, microbiology, computer science, engineering, and materials science in order to create a sensitive and selective manganese sensor that will inform the basic biology of manganese homeostasis, define the manganese metalloproteome, and serve as a device to detect manganese levels in biofluids.

Innovation: This project integrates well established methods from a broad spectrum of disciplines in order to repurpose the high manganese binding affinity of the protein calprotectin into a chemical sensor.

Bioss Team 2 - Workflow

Objective 1
Develop a protein–based Mn2+ specific chelator and determine the microbial response to limiting versus abundant Mn2+ availability

Bioss Team 2 - Figure 1High resolution crystal structure of CP Mn2+ Binding site

Bioss Team 2 - Figure 2D. radidurans, a radiation resistant Organism with an extraordinarily high intracellular Mn2+ Concentration.

Objective 2
Identify proteins that require Mn2+ as a cofactor based on transcriptional profiling. Determine structures of novel Mn-binding proteins using
experimental approaches or comparative modeling.

Bioss Team 2 - Figure 3Schematic of transcriptome profilingBioss Team 2 - Figure 4Crystal structure of Mn superoxide dismutase enzyme

Bioss Team 2 - Figure 5Schematic of microfluidic channels lined With polymer/nanofiber scaffolds with attached protein Mn2+ sensor.

Students Trained:

Graduate Students: Sashari Pinnace, Joshua Haynes, Velia Garcia, Saffron Little

Undergraduate: Isis  Christopher, Anastacia Smith, Rocky Giwa, Kofi Amoah, Emmanuel Jackson, Kinara Byrd, Marshae Lashley, Dana Franklin, Jasmin Jordan, DarleanMartin, RukiayahWarner, Joel Wallace

Nashville State Community College: Horacio Guendulain

Bioss Team 2 - Student 2Bioss Team 2 - Student 1

Bioss Team 2 - Student 3Bioss Team 2 - Student 4

Bioss Team 2