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Protein Footprinting
Protein footprinting is a term used to refer to a method of biochemical analysis that investigates protein structure, assembly, and interactions within a larger macromolecular assembly. It was originally coined in reference to the use of limited proteolysis to investigate contact sites within a monoclonal antibody - protein antigen complex and a year later to examine the protection from hydroxyl radical cleavage conferred by a protein bound to DNA within a DNA-protein complex. In DNA footprinting the protein is envisioned to make an imprint (or footprint) at a particular point of interaction. This latter method was adapted through the direct treatment of proteins and their complexes with hydroxyl radicals and can be generally denoted RP-MS (for Radical Probe - Mass Spectrometry) akin to the designation used for Hydrogen-deuterium exchange Mass Spectrometry (denoted HD-MS or HX-MS). Hydroxyl radical protein footprinting Time-resolved hydroxyl radical protein footprinting (HRPF) emplo ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protein Structure
Protein structure is the three-dimensional arrangement of atoms in an amino acid-chain molecule. Proteins are polymers specifically polypeptides formed from sequences of amino acids, the monomers of the polymer. A single amino acid monomer may also be called a ''residue'' indicating a repeating unit of a polymer. Proteins form by amino acids undergoing condensation reactions, in which the amino acids lose one water molecule per reaction in order to attach to one another with a peptide bond. By convention, a chain under 30 amino acids is often identified as a peptide, rather than a protein. To be able to perform their biological function, proteins fold into one or more specific spatial conformations driven by a number of non-covalent interactions such as hydrogen bonding, ionic interactions, Van der Waals forces, and hydrophobic packing. To understand the functions of proteins at a molecular level, it is often necessary to determine their three-dimensional structure. This is t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protein Folding
Protein folding is the physical process by which a protein chain is translated to its native three-dimensional structure, typically a "folded" conformation by which the protein becomes biologically functional. Via an expeditious and reproducible process, a polypeptide folds into its characteristic three-dimensional structure from a random coil. Each protein exists first as an unfolded polypeptide or random coil after being translated from a sequence of mRNA to a linear chain of amino acids. At this stage the polypeptide lacks any stable (long-lasting) three-dimensional structure (the left hand side of the first figure). As the polypeptide chain is being synthesized by a ribosome, the linear chain begins to fold into its three-dimensional structure. Folding of many proteins begins even during translation of the polypeptide chain. Amino acids interact with each other to produce a well-defined three-dimensional structure, the folded protein (the right hand side of the figure), ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Laboratory Techniques
A laboratory (; ; colloquially lab) is a facility that provides controlled conditions in which scientific or technological research, experiments, and measurement may be performed. Laboratory services are provided in a variety of settings: physicians' offices, clinics, hospitals, and regional and national referral centers. Overview The organisation and contents of laboratories are determined by the differing requirements of the specialists working within. A physics laboratory might contain a particle accelerator or vacuum chamber, while a metallurgy laboratory could have apparatus for casting or refining metals or for testing their strength. A chemist or biologist might use a wet laboratory, while a psychologist's laboratory might be a room with one-way mirrors and hidden cameras in which to observe behavior. In some laboratories, such as those commonly used by computer scientists, computers (sometimes supercomputers) are used for either simulations or the analysis of data. Scienti ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
DNA Profiling
DNA profiling (also called DNA fingerprinting) is the process of determining an individual's DNA characteristics. DNA analysis intended to identify a species, rather than an individual, is called DNA barcoding. DNA profiling is a forensic technique in criminal investigations, comparing criminal suspects' profiles to DNA evidence so as to assess the likelihood of their involvement in the crime. It is also used in paternity testing, to establish immigration eligibility, and in genealogical and medical research. DNA profiling has also been used in the study of animal and plant populations in the fields of zoology, botany, and agriculture. Background Starting in the 1980s, scientific advances allowed the use of DNA as a material for the identification of an individual. The first patent covering the direct use of DNA variation for forensicsUS5593832A was filed by Jeffrey Glassberg in 1983, based upon work he had done while at Rockefeller University in the United States in 1981. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protein Sequencing
Protein sequencing is the practical process of determining the amino acid sequence of all or part of a protein or peptide. This may serve to identify the protein or characterize its post-translational modifications. Typically, partial sequencing of a protein provides sufficient information (one or more sequence tags) to identify it with reference to databases of protein sequences derived from the conceptual translation of genes. The two major direct methods of protein sequencing are mass spectrometry and Edman degradation using a protein sequenator (sequencer). Mass spectrometry methods are now the most widely used for protein sequencing and identification but Edman degradation remains a valuable tool for characterizing a protein's ''N''-terminus. Determining amino acid composition It is often desirable to know the unordered amino acid composition of a protein prior to attempting to find the ordered sequence, as this knowledge can be used to facilitate the discovery of errors i ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Peptide Mass Fingerprinting
Peptide mass fingerprinting (PMF) (also known as protein fingerprinting) is an analytical technique for protein identification in which the unknown protein of interest is first cleaved into smaller peptides, whose absolute masses can be accurately measured with a mass spectrometer such as MALDI-TOF or ESI-TOF. The method was developed in 1993 by several groups independently. The peptide masses are compared to either a database containing known protein sequences or even the genome. This is achieved by using computer programs that translate the known genome of the organism into proteins, then theoretically cut the proteins into peptides, and calculate the absolute masses of the peptides from each protein. They then compare the masses of the peptides of the unknown protein to the theoretical peptide masses of each protein encoded in the genome. The results are statistically analyzed to find the best match. The advantage of this method is that only the masses of the peptides have to be ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Hydrogen–deuterium Exchange
Hydrogen–deuterium exchange (also called H–D or H/D exchange) is a chemical reaction in which a covalently bonded hydrogen atom is replaced by a deuterium atom, or vice versa. It can be applied most easily to exchangeable protons and deuterons, where such a transformation occurs in the presence of a suitable deuterium source, without any catalyst. The use of acid, base or metal catalysts, coupled with conditions of increased temperature and pressure, can facilitate the exchange of non-exchangeable hydrogen atoms, so long as the substrate is robust to the conditions and reagents employed. This often results in perdeuteration: hydrogen-deuterium exchange of all non-exchangeable hydrogen atoms in a molecule. An example of exchangeable protons which are commonly examined in this way are the protons of the amides in the backbone of a protein. The method gives information about the solvent accessibility of various parts of the molecule, and thus the tertiary structure of the protein. T ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
John Wiley & Sons
John Wiley & Sons, Inc., commonly known as Wiley (), is an American multinational publishing company founded in 1807 that focuses on academic publishing and instructional materials. The company produces books, journals, and encyclopedias, in print and electronically, as well as online products and services, training materials, and educational materials for undergraduate, graduate, and continuing education students. History The company was established in 1807 when Charles Wiley opened a print shop in Manhattan. The company was the publisher of 19th century American literary figures like James Fenimore Cooper, Washington Irving, Herman Melville, and Edgar Allan Poe, as well as of legal, religious, and other non-fiction titles. The firm took its current name in 1865. Wiley later shifted its focus to scientific, technical, and engineering subject areas, abandoning its literary interests. Wiley's son John (born in Flatbush, New York, October 4, 1808; died in East Orange, New Je ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Doubly Labeled Water
Doubly labeled water is water in which both the hydrogen and the oxygen have been partly or completely replaced (i.e. ''labeled'') with an uncommon isotope of these elements for tracing purposes. In practice, for both practical and safety reasons, almost all recent applications of the "doubly labeled water" method use water labeled with heavy but non-radioactive forms of each element (deuterium and oxygen-18). In theory, radioactive heavy isotopes of the elements could be used for such labeling; this was the case in many early applications of the method. In particular, doubly labeled water (DLW) can be used for a method to measure the average daily metabolic rate of an organism over a period of time (often also called the Field metabolic rate, or FMR, in non-human animals). This is done by administering a dose of DLW, then measuring the elimination rates of deuterium and oxygen-18 in the subject over time (through regular sampling of heavy isotope concentrations in body water, by ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Flash Photolysis
Flash photolysis is a pump-probe laboratory technique, in which a sample is first excited by a strong pulse of light from a pulsed laser of nanosecond, picosecond, or femtosecond pulse width or by another short-pulse light source such as a flash lamp. This first strong pulse is called the pump pulse and starts a chemical reaction or leads to an increased population for energy levels other than the ground state within a sample of atoms or molecules. Typically the absorption of light by the sample is recorded within short time intervals (by a so-called test or probe pulses) to monitor relaxation or reaction processes initiated by the pump pulse. Flash photolysis was developed shortly after World War II as an outgrowth of attempts by military scientists to build cameras fast enough to photograph missiles in flight. The technique was developed in 1949 by Manfred Eigen, Ronald George Wreyford Norrish and George Porter, who won the 1967 Nobel Prize in Chemistry for this invention. O ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Macromolecular Assembly
The term macromolecular assembly (MA) refers to massive chemical structures such as viruses and non-biologic nanoparticles, cellular organelles and membranes and ribosomes, etc. that are complex mixtures of polypeptide, polynucleotide, polysaccharide or other polymeric macromolecules. They are generally of more than one of these types, and the mixtures are defined spatially (i.e., with regard to their chemical shape), and with regard to their underlying chemical composition and structure. Macromolecules are found in living and nonliving things, and are composed of many hundreds or thousands of atoms held together by covalent bonds; they are often characterized by repeating units (i.e., they are polymers). Assemblies of these can likewise be biologic or non-biologic, though the MA term is more commonly applied in biology, and the term supramolecular assembly is more often applied in non-biologic contexts (e.g., in supramolecular chemistry and nanotechnology). MAs of macromolecules ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Electrical Discharge
An electric discharge is the release and transmission of electricity in an applied electric field through a medium such as a gas (ie., an outgoing flow of electric current through a non-metal medium).American Geophysical Union, National Research Council (U.S.). Geophysics Study Committee (1986) ''The earth's electrical environment''. National Academy Press, Washington, DC, p. 263. Applications The properties and effects of electric discharges are useful over a wide range of magnitudes. Tiny pulses of current are used to detect ionizing radiation in a Geiger–Müller tube. A low steady current can be used to illustrate the spectrum of gases in a gas-filled tube. A neon lamp is an example of a gas-discharge lamp, useful both for illumination and as a voltage regulator. A flashtube generates a short pulse of intense light useful for photography by sending a heavy current through a gas arc discharge. Corona discharges are used in photocopiers. Electric discharges can convey substa ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
