The Relation of Structure to Enzymatic Activity in Ribonuclease*

Kalnitsky, George; Hummel, John P.; Resnick, Harold; Carter, J. R.; Barnett, Lewis B.; Dierks, Christa

doi:10.1111/j.1749-6632.1959.tb49336.x

Cited by 91 publications

(7 citation statements)

References 32 publications

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“…The nal protein concentration was determined by measuring absorbance of protein at 278 nm using a Thermo Scientic Helios Zeta double-beam UV/VIS spectrophotometer (RNase A has an extinction coefficient of 8640 cm À1 M À1 provided by the manufacturer). The activity of the dialyzed enzyme was checked using the method of Kalnitsky et al 52 DSC experiments were performed using a Nano DSC (TA Instruments) with 0.3 mL capillary cell volume. Samples were equilibrated at 20 C and then heated to 85 C with a scan rate of 1 C per minute and at 3 atm.…”

Section: Differential Scanning Calorimetry (Dsc) Measurementsmentioning

confidence: 99%

Do soft anions promote protein denaturation through binding interactions? A case study using ribonuclease A

et al. 2019

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Section: Differential Scanning Calorimetry (Dsc) Measurementsmentioning

confidence: 99%

Do soft anions promote protein denaturation through binding interactions? A case study using ribonuclease A

et al. 2019

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“…35 For this purpose, free RNase A and AuNPs-PEG-RNase A conjugates were loaded on 12% polyacrylamide gel and then protein bands were visualized by staining with coomassie brilliant blue G-250. Besides, the RNase A activity in free and conjugated form was determined by measuring the RNA degradation using the method of Kalnitsky et al 36 Briefly, the solution of 1 mg/mL of the enzyme in 0.10 M sodium acetate was prepared (pH 5.0). Afterward, 1 mL of enzyme solution was added to different concentrations of 1% RNA (0.5, 1, 2, 5 and 10 μg/mL) and then all samples were incubated at 37°C for 5 minutes.…”

Section: Confirmation Of Aunps-peg-rnase a Conjugates And Enzyme Actimentioning

confidence: 99%

PEGylated gold nanoparticles-ribonuclease induced oxidative stress and apoptosis in colorectal cancer cells

et al. 2019

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Introduction: Currently, drug-induced reactive oxygen species (ROS) mediating apoptosis pathway have extensively been investigated in designing effective strategies for colorectal cancer (CRC) chemotherapy. Bovine pancreatic ribonuclease A (RNase A) represents a new class of cytotoxic and non-mutagenic enzymes, and has gained more attention as a potential anticancer modality; however, the cytosolic ribonuclease inhibitors (RIs) restrict the clinical application of this enzyme. Nowadays, nanotechnology-based diagnostic and therapeutic systems have provided potential solutions for cancer treatments. Methods: In this study, the gold nanoparticles (AuNPs) were synthesized, stabilized by polyethylene glycol (PEG), functionalized, and covalently conjugated with RNase A. The physicochemical properties of engineered nanobiomedicine (AuNPs-PEG-RNase A) were characterized by scanning electron microscope (SEM), dynamic light scattering (DLS), and UV-vis spectrum. Then, its biological impacts including cell viability, apoptosis, and ROS production were evaluated in the SW-480 cells. Results: The engineered nanobiomedicine, AuNPs-PEG-RNase A, was found to effectively induce apoptosis in SW-480 cells and result in a significant reduction in cancer cell viability. Besides, the maximum production of ROS was obtained after the treatment of cells with an IC50 dose of AuNPs-PEG-RNase A. Conclusion: Based on the efficient ROS-responsiveness and the anticancer activity of RNase A of the engineered nanomedicine, this nanoscaled biologics may be considered as a potential candidate for the treatment of CRC.

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“…This refers to his previous (wrong) conclusion that an ordered shape of a protein is not strictly needed for its catalytic function if the structure of the active site is intact. In fact, further studies revealed that some apparent increment of activity occurs at high urea concentration probably due to an increased solubility of the reaction product or to a denaturation of the RNA thereby making it more available to the digestion by the enzyme [ 25 ]. In reality, the RNase activity is slightly lowered at similar urea concentration [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

The Anfinsen Dogma: Intriguing Details Sixty-Five Years Later

Gambardella

Notari

Cavaterra

et al. 2022

IJMS

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The pioneering experiments of Anfinsen on the oxidative folding of RNase have been revisited discovering some details, which update the statement of his dogma and shed new light on the leading role of the correct disulfide in the attainment of the native structure. CD analysis, mass spectrometry, fluorescence spectroscopy and enzyme activity indicate that native disulfides drive the formation of the secondary and tertiary structures that cannot be entirely formed in their absence. This opposes a common opinion that these structures are first formed and then stabilized by the native disulfides. Our results also indicate that a spontaneous re-oxidation of a reduced RNase cannot produce a complete recovery of activity, as described by many textbooks; this can be obtained only in the presence of a reshuffling solution such as GSH/GSSG.

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The Relation of Structure to Enzymatic Activity in Ribonuclease*

Cited by 91 publications

References 32 publications

Do soft anions promote protein denaturation through binding interactions? A case study using ribonuclease A

Do soft anions promote protein denaturation through binding interactions? A case study using ribonuclease A

PEGylated gold nanoparticles-ribonuclease induced oxidative stress and apoptosis in colorectal cancer cells

The Anfinsen Dogma: Intriguing Details Sixty-Five Years Later

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