tailieunhanh - Báo cáo khoa học: A fluorescence energy transfer-based mechanical stress sensor for specific proteins in situ

To measure mechanical stress in real time, we designed a fluorescence reso-nance energy transfer (FRET) cassette, denoted stFRET, which could be inserted into structural protein hosts. The probe was composed of a green fluorescence protein pair, Cerulean and Venus, linked with a stablea-helix. | ễFEBS Journal A fluorescence energy transfer-based mechanical stress sensor for specific proteins in situ Fanjie Meng Thomas M. Suchyna and Frederick Sachs Center for Single Molecule Biophysics Department of Physiology and Biophysics State University of New York at Buffalo NY USA Keywords Cerulean fluorescence resonance energy transfer relative orientation factor Venus a-helix linker Correspondence F. Sachs Center for Single Molecule Biophysics Department of Physiology and Biophysics State University of New York at Buffalo 3435 Main Street Buffalo NY 14214 USA Fax 1 716 829 2569 Tel 1 716 829 3289 ext. 105 E-mail sachs@ Received 15 December 2007 revised 9 April 2008 accepted 11 April 2008 doi To measure mechanical stress in real time we designed a fluorescence resonance energy transfer FRET cassette denoted stFRET which could be inserted into structural protein hosts. The probe was composed of a green fluorescence protein pair Cerulean and Venus linked with a stable a-helix. We measured the FRET efficiency of the free cassette protein as a function of the length of the linker the angles of the fluorophores temperature and urea denaturation and protease treatment. The linking helix was stable to 80 C unfolded in 8 M urea and rapidly digested by proteases but in all cases the fluorophores were unaffected. We modified the a-helix linker by adding and subtracting residues to vary the angles and distance between the donor and acceptor and assuming that the cassette was a rigid body we calculated its geometry. We tested the strain sensitivity of stFRET by linking both ends to a rubber sheet subjected to equibiaxial stretch. FRET decreased proportionally to the substrate strain. The naked cassette expressed well in human embryonic kidney-293 cells and surprisingly was concentrated in the nucleus. However when the cassette was located into host proteins such a-actinin nonerythrocyte spectrin and filamin A the labeled hosts .

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• Crystal structure
• medical analysis
• biochemical studies
• Zizyphus mauritiana Lamk
• Biochemical constitutes
• Ber selection 1
• Studies on the pattern
• Pattern of changes biochemical constitutes
• Studies on Tikka disease
• Tikka disease
• Sowing dates
• Biochemical parameters of groundnut
• Biochemical parameters
• Studies on biochemical changes
• Biochemical changes
• Cell wall degrading enzymes
• Papaya fruit inoculated
• Colletotrichum demetium
• Vindhyan zone
• Haemato biochemical profile
• Haemato biochemical profile of goats
• Goats in Vindhyan region
• Biochemical variables
• Diaphragmatic hernia
• Various NSAIDS protocols affected
• Buffaloes treated
• Biochemical composition
• Reducing and Non reducing sugar
• Non reducing sugar
• Biochemical composition of various tomato
• Various tomato (Solanum lycopersicum L )
• environmental medicine
• Hepatobiliary disorders
• Serum biochemistry
• Hemato biochemical studies
• Hepatobiliary disorders in dogs
• Elevated total leucocyte
• Agricultural science
• Physiological and biochemical studies
• Wheat plants by application
• Amino acids and yeast extract
• Physio chemical
• Water relations
• Lucknow 49
• Ascarbic acid
• Different cultivars
• Guava under eastern
• Biochemical studies on adverse health
• Adverse health effects of arsenic
• Chromium in buffaloes
• Level of arsenic and chromium
• Heavy metals in buffaloes
• PyometraCEH complex
• Haemato biochemical evaluation
• Bitches affected
• Cystic endometrial hyperplasia pyometra complex
• Haemato biochemical studies
• Sloth bears
• HematoBiochemical changes
• Biochemical values
• Hemato biochemical changes
• Xylazine Zolazepam
• Sahiwal cows
• Blood metabolites
• Complete lactation in Sahiwal cows
• breast cancer
• medicine
• Hematopoietic
• medical knowledge