Sodium dodecyl sulfate polyacrylamide gel electrophoresis also known as SDS-PAGE is one of the methods for determining the molecular weight of unknown proteins. SDS is an anionic molecule which denaturizes proteins and brings it back to its’ primary structure and it also provides a negative charge to the uncharged molecule. The SDS-PAGE enables the separation of proteins based on their sizes. The larger the size of the protein, the harder it is to travel through the gel thus heavier proteins stay near the cathode side of the gel. For this experiment, a software named Gel Analyzer was used in order to obtain the molecular weight of the unknown proteins with the help of a protein ladder with known molecular weight and protein concentration.
However, all proteins are constructed from the same set of 20 amino acids linked in unbranched polymers. The covalent bond that exists between amino acids is called peptide bond, hence a polymer of amino acids is named polypeptide. A protein is a biological functional molecule made up of one or more polypeptides which is folded and coiled into unique three-dimensional structure. In laboratory, it is important to measure the concentration of proteins for research investigations. Biuret test is adopted to quantify proteins in fluid by using a spectrophotometer.
Amino acids are the building blocks of proteins. All amino acids have the same basic structure but differ in their R-side chains. Each amino acid consists of an amino group (-NH3), a carboxyl group (-COOH) and a hydrogen atom (H). The amino and carboxyl groups are attached to a central alpha carbon together with a hydrogen atom and an R-side chain. There are currently known that over 170 amino acids occur in organisms but only 20 are commonly found in proteins.
Objective: The purpose of this experiment is to determine the changes in activity level (if any) when enzymes are exposed to a variety of environments (in this case, temperature). Introduction: Enzymes are made up of a series of proteins known as amino acids. Enzymes are essential in almost all aspects of life. There are certain ways to identify an enzyme by the name for each, for example they usually end with the suffix -ase. For instance, amylase (present in saliva), lipase, or protease.
ABSTRACT Gamma secretase enzyme is a multi-subunit proteinase complex, an integral membrane protein that severs single-pass transmembrane proteins at residues intervals the transmembrane domain. The most substrates of γ- secretase are amyloid precursor protein (APP), an outsized integral membrane macromolecule that, once cleaved by each γ-and β-secretase, produces 39-42 amino acid amide known as amyloid beta whose abnormally folded fibrillar type is that the primary part of amyloid plaques found in the brains of Alzheimer's disease (AD) patients. The gamma-secretase complex consists of 4 individual proteins: presenilin, PEN-2 (presenilin enhancer 2), APH-1 (anterior pharynx-defective 1), nicastrin. We know the structure of 2 subunits presenilin and
It may limit the application of the PS cyclization as a chemical ligation method for peptides with N-terminal aromatic residue and peptides with aldehyde residue at C-terminal(40). 7. Pictet-spengler reaction for protein chemical modification= proteins are having aldehyde and ketone groups in their structures. So proteins are taken as a substrate and the pictet-spengler reaction is performed for making modification in the chemical nature of the proteins. P. agarwal and co-workers work for protein chemical modification by conducting a pictet-spengler reaction between aldehydes and alkoxyamines.
Biosynthesis Pathway A biosynthesis pathway describes the steps that take place in a chemical reaction which occurs when living organisms create new molecule from simpler ones. The word "biosynthesis" comes from two words: "bio," which means that the reaction is occurring in living organism and "synthesis," which indicates that large products are made up by simpler molecules. To describe a pathway completely some compounds are involved which includes such as which enzymes, coenzymes and cofactors are used in each reaction. Not all molecules are synthesized by humans, some molecules such as some essential amino acids i.e. lysine, these nutrients come from the protein rich food we consume including beans.
KINETICS OF MULTISUBSTRATE REACTIONS Introduction Enzyme kinetics is the study of rate of biochemical reactions that are catalyzed by enzymes. In enzyme kinetics, the reaction rate is measured and the their effect is measured or investigated. Studying an enzyme kinetics in this way we can check the catalytic activity of enzyme, its major role in metabolism, and how its activity is determined. Enzymes are protein in nature and binds to substrates. These substrate molecules bind to active site of enzyme and changed into products through a number of steps known as enzymatic reactions.
1.Introduction: An enzyme is a large protein that acts as a biological catalyst which changes the rate of a reaction. It provides an active site which is an environment where a reaction can take place this is made up of amino acids. The structure and shape of the substrate, the structure and shape of an enzyme and the substance upon which the enzyme works all have to match exactly. This enables the substrate to bind, but it can 't do this if the shapes of the two are different. The Aim of Enzyme Catalase Experiment is making a series of experiments involving the enzyme Catalase which has been performed in order to determine some of the enzyme 's properties.