- ELSEVIER SCIENCE BV
- Biblioteca Digitais de Teses e Dissertações da USP
- Universidade Estadual Paulista
- Biblioteca Digital da Unicamp
- The Biophysical Society
- Universidade Federal de Goiás; BR; UFG; Mestrado em Nutricao e Saude; Ciencias da Saude
- Universidade de Adelaide
- Universidade de Tubinga
- La Sapienza Universidade de Roma
- Quens University
- Universidade Duke
- University of Delaware
- Nutrición Hospitalaria
- South African Journal of Animal Science
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Effects of dietary urea levels on milk protein fractions of Holstein cows
Suplementação com aminoácios de cadeia ramificada atenua em proles os efeitos mediados pela dieta materna restrita em proteína; Branched-chain amino acids supplementation attenuates in offspring the effects mediated by maternal protein-restrict diet.
Evaluation of a protein deficient diet in rats through blood oxidative stress biomarkers
Experimental mixture design as a tool for proteases production by Aspergillus niger and obtaining of protein hydrolysates with multiple functional and biological properties; Aplicação da ferramenta de planejamento experimental de misturas como estratégia para a produção de proteases por Aspergillus niger e obtenção de hidrolisados proteicos com múltiplas propriedades funcionais e biológicas
Statistics and Physical Origins of pK and Ionization State Changes upon Protein-Ligand Binding
Eficácia alimentar e qualidade proteica de misturas de caseína com gelatina em dietas com baixos teores de proteína; Food and protein efficiency mixtures of gelatin with casein in diets with low levels of protein
The effect of higher protein human milk fortifier on growth in preterm infants.
Parallel and miniaturised Analysis of Protein-Protein Interactions in T-Cell Signal Transduction by Fluorescence Cross-Correlation Spectroscopy and Peptide Microarrays; Parallele und miniaturisierte Analyse von Protein-Protein-Interaktionen in der T-Zell-Signaltransduktion mittels Fluoreszenz-Kreuzkorrelations-Spektroskopie und Peptidmikroarrays
Funktionelle Untersuchungen von Colicin M und seinem Immunitätsprotein; Functional analysis of colicin M and its immunity protein
Modeling Flexibility of Protein-DNA and Protein-Ligand Complexes using Molecular Dynamics; Modellierung der Flexibilität von Protein-DNA und Protein-Ligand Komplexen mit Hilfe von Molekulardynamischen Simulationen
Resource for benchmarking the applicability of protein structure models
Pattern Discovery in Protein Structures and Interaction Networks
Development and Application of a quantitative Mass spectrometry based Platform for Thermodynamic Analysis of Protein interaction Networks
The identification and quantification of protein-protein interactions in large scale is critical to understanding biological processes at a systems level. Current approaches for the analysis of protein -protein interactions are generally not quantitative and largely limited to certain types of interactions such as binary and strong binding interactions. They also have high false-positive and false-negative rates. Described here is the development of and application of mass spectrometry-based proteomics metehods to detect and quantify the strength of protein-protein and protein-ligand interactions in the context of their interaction networks. Characterization of protein-protein and protein-ligand interactions can directly benefit diseased state analyses and drug discovery efforts.
The methodologies and protocols developed and applied in this work are all related to the Stability of Unpurified Proteins from Rates of amide H/D Exchange (SUPREX) and Stability of Protein from Rates of Oxidation (SPROX) techniques, which have been previously established for the thermodynamic analysis of protein folding reactions and protein-ligand binding interactions. The work in this thesis is comprised of four parts. Part I involves the development of a Histidine Slow H/D exchange protocol to facility SURPEX-like measurements on the proteomic scale. The Histidine Slow H/D exchange protocol is developed in the context of selected model protein systems and used to investigate the thermodynamic properties of proteins in a yeast cell lysate.
In Part II an isobaric mass tagging strategy is used in combination with SPROX (i.e....
Development and Applications of Chemical Labeling Protocols for Protein-Ligand Binding Analysis Using Bottom-Up Proteomics
Proteins fold into well-defined three-dimensional structures to carry out their biological functions which involve non-covalent interactions with other cellular molecules. Knowledge about the thermodynamic properties of proteins and protein-ligand complexes is essential for answering fundamental biological questions and drug or biomarker discovery. Recently, chemical labeling strategies have been combined with mass spectrometry methods to generate thermodynamic information about protein folding and ligand binding interactions. The work in this thesis is focused on the development and application of two such chemical labeling protocols coupled with mass spectrometry including one termed, SUPREX (stability of unpurified proteins from rates of H/D exchange), and one termed SPROX (stability of proteins from rates of oxidation). The work described in this thesis is divided into two parts. The first part involves the application of SUPREX to the thermodynamic analysis of a protein folding chaperone, Hsp33, and its interaction with unfolded protein substrates. The second part involves the development of a new chemical labeling protocol that can be used to make protein folding and ligand binding measurements on the proteomic scale.
In the first part of this work...
Novel Algorithms for Computational Protein Design, with Applications to Enzyme Redesign and Small-Molecule Inhibitor Design
Computational protein design aims at identifying protein mutations and conformations with desired target properties (such as increased protein stability, switch of substrate specificity, or novel function) from a vast combinatorial space of candidate solutions. The development of algorithms to efficiently and accurately solve problems in protein design has thus posed significant computational and modeling challenges. Despite the inherent hardness of protein design, a number of computational techniques have been previously developed and applied to a wide range of protein design problems. In many cases, however, the available computational protein design techniques are deficient both in computational power and modeling accuracy. Typical simplifying modeling assumptions for computational protein design are the rigidity of the protein backbone and the discretization of the protein side-chain conformations. Here, we present the derivation, proofs of correctness and complexity, implementation, and application of novel algorithms for computational protein design that, unlike previous approaches, have provably-accurate guarantees even when backbone or continuous side-chain flexibility are incorporated into the model. We also describe novel divide-and-conquer and dynamic programming algorithms for improved computational efficiency that are shown to result in speed-ups of up to several orders of magnitude as compared to previously-available techniques. Our novel algorithms are further incorporated as part of K*...
Kinetic Characterization of the Coupled Folding and Binding Mechanism of Bacterial RNase P Protein: an Intrinsically Unstructured Protein
Understanding the interconversion between the thermodynamically distinguishable states present in a protein folding pathway provides not only the kinetics and energetics of protein folding but also insights into the functional roles of these states in biological systems. The protein component of bacterial RNase P holoenzyme from Bacillus subtilis (P protein) was used as a model system to elucidate the general folding/unfolding of an intrinsically unstructured protein (IUP) both in the absence and presence of ligands.
P protein was previously characterized as an intrinsically unstructured protein, and it is predominantly unfolded in the absence of ligands. Addition of small anions can induce the protein to fold. Therefore, the folding and binding are tightly coupled. Trimethylamine-N oxide (TMAO), an osmolyte that stabilizes the unliganded folded form of the protein, enabled us to study the folding process of P protein in the absence of ligand. Transient stopped-flow kinetic time courses at various final TMAO concentrations showed multiphase kinetics. Equilibrium "cotitration" experiments were performed using both TMAO and urea to obtain a TMAO-urea titration surface of P protein. Both kinetic and equilibrium studies show evidence of an intermediate state in the P protein folding process. The intermediate state is significantly populated and the folding rate constants involved in the reaction are slow relative to similar size proteins.
NMR spectroscopy was used to characterize the structural properties of the folding intermediate of P protein. The results indicate that the N-terminal (residues 2-19) and C-terminal regions (residues 91-116...
Analysis and Redesign of Protein-Protein Interactions: A Hotspot-Centric View
One of the most significant discoveries from mutational analysis of protein interfaces is that often a large percentage of interface residues negligibly perturb the binding energy upon mutation, while residues in a few critical "hotspots" drastically reduce affinity when mutated. The organization of protein interfaces into hotspots has a number of important implications. For example, small interfaces can have high affinity, and when multiple binding partners are generated to the same protein, they are predisposed to binding the same regions and often have the same hotspots. Even small molecules that bind to interfaces and disrupt protein-protein interactions (PPIs) tend to bind at hotspots. This suggests that some hotspot-forming sites on protein surfaces are
In order to gain insight into the nature of hotspots I experimentally examined the small, but high-affinity interface between the synthetically evolved ankyrin repeat protein Off7 with E. coli maltose binding protein by characterization of mutant variants and redesigned interfaces. In order to characterize many mutants...