scholarly journals Structural lesion to ribonuclease A caused by reductive stress: Assessment by Raman spectroscopy

2008 ◽  
Vol 22 (4) ◽  
pp. 279-286 ◽  
Author(s):  
Armida Torreggiani
Keyword(s):  
Raman Spectroscopy ◽  
Conformational Changes ◽  
Protein Secondary Structure ◽  
Disulfide Bridge ◽  
Acid Sites ◽  
Ribonuclease A ◽  
Rnase A ◽  
Structural Lesion ◽  
Pancreatic Ribonuclease A ◽  
Sulfur Containing

The damages induced by reductive radical stress on bovine pancreatic ribonuclease A (RNAse A) were investigated by Raman spectroscopy. Gamma-irradiation was used to simulate the endogenous formation of reductive species, in particular•H atom that is a simple one-electron equivalent reducing agent. Specific damages occur at sensitive amino acid sites, selectively, rather than indiscriminately, leading to the structure modification of the protein. Sulfur-containing residues (Met and Cys) and aromatic residues are appreciably attacked. In particular, extensive changes in the disulfide bridge conformations are induced as well as conformational changes of the protein secondary structure; a gradual conversion ofα-helical to pleated-sheet geometry was evidenced, indicating a higher ability of reducing radicals in denaturing the protein structure compared with that of oxidizing radical species.

scholarly journals Amino acids and peptides. XIV Synthesis and biological activity of three S-peptide analogues of bovine pancreatic ribonuclease A (RNase A).

1987 ◽  
Vol 35 (2) ◽  
pp. 468-478 ◽  
Author(s):  
NAOKI TENO ◽  
SATOSHI TSUBOI ◽  
TOMOKO SHIMAMURA ◽  
YOSHIO OKADA ◽  
YOSHIMI YANAGIDA ◽  
...  
Keyword(s):  
Amino Acids ◽  
Biological Activity ◽  
Ribonuclease A ◽  
Rnase A ◽  
Pancreatic Ribonuclease ◽  
Pancreatic Ribonuclease A ◽  
Peptide Analogues

scholarly journals Preliminary investigation of deoxyoligonucleotide binding to ribonuclease A using mass spectrometry: An attempt to develop a lab experience for undergraduates

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 340
Author(s):  
Daniel D. Clark
Keyword(s):  
Mass Spectrometry ◽  
Ion Trap ◽  
Dissociation Constants ◽  
Preliminary Investigation ◽  
Accurate Determination ◽  
Ribonuclease A ◽  
Rnase A ◽  
Protein Ligand Interactions ◽  
Ligand Interactions ◽  
Pancreatic Ribonuclease A

Deoxyoligonucleotide binding to bovine pancreatic ribonuclease A (RNase A) was investigated using electrospray ionization ion-trap mass spectrometry (ESI-IT-MS). Deoxyoligonucleotides included CCCCC (dC5) and CCACC (dC2AC2).  This work was an attempt to develop a biochemistry lab experience that would introduce undergraduates to the use of mass spectrometry for the analysis of protein-ligand interactions.  Titration experiments were performed using a fixed RNase A concentration and variable deoxyoligonucleotide concentrations.  Samples at equilibrium were infused directly into the mass spectrometer under native conditions.  For each deoxyoligonucleotide, mass spectra showed one-to-one binding stoichiometry, with marked increases in the total ion abundance of ligand-bound RNase A complexes as a function of concentration, but the accurate determination of dC5 and dC2AC2 dissociation constants was problematic.

Influence of vanadium(V) complexes on the catalytic activity of ribonuclease A. The role of vanadate complexes as transition state analogues to reactions at phosphate

1996 ◽  
Vol 74 (1) ◽  
pp. 38-48 ◽  
Author(s):  
Chui Har Leon-Lai ◽  
Michael J. Gresser ◽  
Alan S. Tracey
Keyword(s):  
Transition State ◽  
Binding Constants ◽  
Kinetic Studies ◽  
Ribonuclease A ◽  
Rnase A ◽  
Binding Energies ◽  
Phosphoglycerate Mutase ◽  
Binding Studies ◽  
Pancreatic Ribonuclease A

The interactions of vanadate and its complexes of uridine, 5,6-dihydrouridine, and methyl β-D-ribofuranoside with bovine pancreatic ribonuclease A (RNase A) (EC 3.1.27.5) were studied by 51V NMR spectroscopy and enzyme kinetics. From kinetic studies, it was found that neither inorganic vanadate nor the methyl β-D-ribofuranoside–vanadate complex significantly inhibited the RNase A catalyzed hydrolysis of uridine 2′,3′-cyclic monophosphate. The NMR binding studies were in full agreement with the kinetics studies and showed that neither inorganic vanadate nor the methyl β-D-ribofuranoside–vanadate complex was bound tightly by the enzyme. Approximate binding constants were (5.0 ± 1.0) × 10−7 M and (3.0 ± 0.6) × 10−6 M for the uridine–and 5,6-dihydrouridine–vanadate complexes, respectively. An induced-fit mechanism is suggested, in which the pyrimidine subsite of the active site of RNase A must be fully occupied for the enzyme to be able to tightly bind the transition state or transition state analog. Calculation of the binding energies of vanadate complexes in ribonuclease, phosphoglycerate mutase, and phosphoglucomutase revealed an excess of binding energy over the analogous phosphate derivative of about 25 kJ/mol for all enzymes, even though the binding constants themselves varied by about six orders of magnitude. This energy represents about 40% of that expected to be available for a trigonal-bipyramidal transition state and requires a reassessment of the role of vanadate as a transition state analogue for phosphate transfer. Key words: vanadate, ribonuclease, transition state, binding constants, phosphate analogues, kinetics.

scholarly journals Preliminary investigation of deoxyoligonucleotide binding to ribonuclease A using mass spectrometry: An attempt to develop a lab experience for undergraduates

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 340
Author(s):  
Daniel D. Clark
Keyword(s):  
Mass Spectrometry ◽  
Ion Trap ◽  
Dissociation Constants ◽  
Preliminary Investigation ◽  
Accurate Determination ◽  
Ribonuclease A ◽  
Rnase A ◽  
Protein Ligand Interactions ◽  
Ligand Interactions ◽  
Pancreatic Ribonuclease A

Deoxyoligonucleotide binding to bovine pancreatic ribonuclease A (RNase A) was investigated using electrospray ionization ion-trap mass spectrometry (ESI-IT-MS). Deoxyoligonucleotides included CCCCC (dC5) and CCACC (dC2AC2).  This work was an attempt to develop a biochemistry lab experience that would introduce undergraduates to the use of mass spectrometry for the analysis of protein-ligand interactions.  Titration experiments were performed using a fixed RNase A concentration and variable deoxyoligonucleotide concentrations.  Samples at equilibrium were infused directly into the mass spectrometer under native conditions.  For each deoxyoligonucleotide, mass spectra showed one-to-one binding stoichiometry, with marked increases in the total ion abundance of ligand-bound RNase A complexes as a function of concentration, but the accurate determination of dC5 and dC2AC2 dissociation constants was problematic.

scholarly journals On the Preparation of Bovine Pancreatic Ribonuclease A

1963 ◽  
Vol 238 (2) ◽  
pp. 618-621 ◽  
Author(s):  
Arthur M. Crestfield ◽  
William H. Stein ◽  
Stanford Moore
Keyword(s):  
Ribonuclease A ◽  
Pancreatic Ribonuclease ◽  
Pancreatic Ribonuclease A

scholarly journals Structural and Technological Approach to Reveal the Role of the Lipid Phase in the Formation of Soy Emulsion Gels with Chia Oil

Gels ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 48
Author(s):  
Ana M. Herrero ◽  
Claudia Ruiz-Capillas
Keyword(s):  
Raman Spectroscopy ◽  
Structural Properties ◽  
Structural Changes ◽  
Protein Secondary Structure ◽  
Lipid Phase ◽  
Α Helix ◽  
Β Sheet ◽  
Shed Light ◽  
Chia Oil

Considerable attention has been paid to emulsion gels (EGs) in recent years due to their interesting applications in food. The aim of this work is to shed light on the role played by chia oil in the technological and structural properties of EGs made from soy protein isolates (SPI) and alginate. Two systems were studied: oil-free SPI gels (SPI/G) and the corresponding SPI EGs (SPI/EG) that contain chia oil. The proximate composition, technological properties (syneresis, pH, color and texture) and structural properties using Raman spectroscopy were determined for SPI/G and SPI/EG. No noticeable (p > 0.05) syneresis was observed in either sample. The pH values were similar (p > 0.05) for SPI/G and SPI/EG, but their texture and color differed significantly depending on the presence of chia oil. SPI/EG featured significantly lower redness and more lightness and yellowness and exhibited greater puncture and gel strengths than SPI/G. Raman spectroscopy revealed significant changes in the protein secondary structure, i.e., higher (p < 0.05) α-helix and lower (p < 0.05) β-sheet, turn and unordered structures, after the incorporation of chia oil to form the corresponding SPI/EG. Apparently, there is a correlation between these structural changes and the textural modifications observed.

Sign in / Sign up

Export Citation Format

Share Document