The goal of this administrative supplement of the parent award, R35GM118090, is to purchase a Prometheus Panta from NANOTEMPER for the simultaneous detection of nano-Differential Scanning Fluorimetry (nanoDSF), Backreflection, Dynamic Light Scattering (DLS) and Static Light Scattering (SLS). This instrument overcomes many of the drawbacks of the current biophysical technologies available in the lab [Size Exclusion Chromatography (SEC), Analytical Ultracentrifugation (AUC), Mass Photometry (MP), and Differential Scanning Fluorimetry (DSF)], and will provide entirely new capabilities. The Prometheus Panta, requested in this administrative supplement, will allow us to carry out protein and protein complex analysis rapidly, accurately, label-free, and with low sample requirement of ~10 L at a wide range of protein and protein complex concentrations of between 5 µg/mL – 250 mg/ml. This analysis will include protein and protein complex quality assessment and buffer condition optimization for biochemical and structural (X-ray crystallography and single particle cryo-EM) analysis, assessing thermal stability of protein, protein-ligand, protein complexes and protein mutants as a readout for protein ligand binding and protein integrity, respectively. Importantly, this technology will also provide entirely new capabilities to the lab including analysis of samples in the presence of detergents and organic solvents, assessing molecular mass determinations at a wide range of sample concentrations, and carrying out protein aggregation studies. The instrument also has flexible throughput (24 samples using capillary chips or 48 samples when individual capillaries are loaded). The instruments combined technologies comprised of nanoDSF, Backreflection, DLS, and SLS allow the determination of key parameters including melting point (Tm), onset of unfolding temperature (Ton), turbidity formation (Tturb), hydrodynamic radius (rH), self-interaction parameter (kD), polydispersity index (PDI), molecular weight, and second virial coefficient (B22). Obtaining a Prometheus Panta will significantly advance the goals and overall impact of the parent grant. Specific projects that will benefit from the instrumentation include studies on multiprotein protein and RNA acetyltransferases (Sas2/4/5, Caf1 and Nat10), N-terminal protein acetyltransferase (NAT) regulation (NAT complexes and ribosomes), small molecule acetyl-transfer enzymes (ACLY, ACSS2, FASN, ACC1), inhibitor development (NatA, NatB, NatD, ACSS2, ACLY, FASN) and new projects on membrane associated proteins (AANAT and HSD3B7). There is currently no comparable instrument on campus or in the greater Philadelphia area. The instrument will be located in a dedicated biophysical equipment room within the Marmorstein laboratory and Dr. Marmorstein’s home Department of Biochemistry and Biophysics is committed to the proper installation and maintenance of the Mass Photometer throughout its lifetime. Dr Marmorstein will also pro...