Effects of Recombinant Protein Expression on Green Fluorescent Protein Diffusion in Escherichia coli

• Published in: Biochemistry (Easton) Vol. 48; no. 23; pp. 5083 - 5089
• Main Authors: Slade, Kristin M, Baker, Rachael, Chua, Michael, Thompson, Nancy L, Pielak, Gary J
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 16.06.2009
• Subjects: alpha-Synuclein - genetics; alpha-Synuclein - metabolism; Calmodulin - genetics; Calmodulin - metabolism; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cytoplasm - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; Fluorescence Recovery After Photobleaching; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Maltose-Binding Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; tau Proteins - genetics; tau Proteins - metabolism
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi9004107

Fluorescence recovery after photobleaching was used to measure the diffusion coefficient of green fluorescent protein (GFP, 27 kDa) in Escherichia coli in the presence or absence of four coexpressed proteins: cytoplasmic maltose binding protein (42 kDa), tau-40 (45 kDa), α-synuclein (14 kDa), or calmodulin (17 kDa). The GFP diffusion coefficient remains constant regardless of the type of coexpresseed protein and whether or not the coexpressed protein was induced. We conclude that expression of these soluble proteins has little to no effect on the diffusion of GFP. These results have implications for the utility of in-cell nuclear magnetic resonance spectroscopy.