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Proteins engineering

Protein engineering encompasses a vast amount and wide variety of research. At least two textbooks (1,2) have been devoted exclusively to this topic, and several excellent reviews have been pubHshed (3,4). Herein, an overview of principles, an introduction to basic techniques, and a summary of results of representative experiments on protein engineering are provided. [Pg.194]

Much of protein engineering concerns attempts to explore the relationship between protein stmcture and function. Proteins are polymers of amino acids (qv), which have general stmcture +H3N—CHR—COO , where R, the amino acid side chain, determines the unique identity and hence the stmcture and reactivity of the amino acid (Fig. 1, Table 1). Formation of a polypeptide or protein from the constituent amino acids involves the condensation of the amino-nitrogen of one residue to the carboxylate-carbon of another residue to form an amide, also called peptide, bond and water. The linear order in which amino acids are linked in the protein is called the primary stmcture of the protein or, more commonly, the amino acid sequence. Only 20 amino acid stmctures are used commonly in the cellular biosynthesis of proteins (qv). [Pg.194]

Kirk-Othmer Encyclopedia of Chemical Technology (4th Edition) [Pg.194]

Amino acid Three- letter code One- letter code Mass of residue in. b proteins Accessible surface area, 2 nm Hydrophobicity index ionizable side chain Occurrence in n/ proteins, /o Relative mutabihty [Pg.195]

Values reflect the moleculai weights of the amino acids minus that of water. [Pg.195]

Protein engineering is a hybrid discipline wherein recombinant DNA technology, conventional protein chemistry and a host of biochemical, chemical and physical techniques are applied to design, produce and investigate proteins with two purposes. [Pg.501]

dissection of the stracture and activity of existing proteins by making systematic alternations of their stractures and examining the changes in their function and properties and [Pg.501]

production of novel proteins de novo or altered proteins to give useful changes in activities and/or properties. [Pg.501]

Two classes of functional proteins will be considered. This snbsection considers engineering enzymes and the next subsection deals with engineering antibodies. [Pg.501]

The ideal mutation for enzyme studies is a nondisruptive deletion, i.e. only ranov-ing an interaction without causing a dismption or reorganization of protein structure. The alternations (mutations) causing least structural effect are  [Pg.501]

The following sections discuss the application of HPLC to the various methods used in the isolation and analysis of proteins, peptides and amino acids. The development of new stationary phases coupled with instrumentation which allows unattended gradient development has transformed the task of purification. Some of these aspects will also be discussed in this section. [Pg.172]

Ion-exchange chromatography (lEC) is the most widely used mode for the separation of proteins, since the optimum chromatographic conditions are compatible with the maintenance of the secondary and tertiary structure of complex biopolymers. The retention of a solute in lEC can be controlled by altering the ionic strength, pH and temperature. In addition it has been observed that these parameters can also [Pg.172]

The stationary phases used for lEC of proteins are comprised of spherical beads which possess a large pore size (30-50 nm). The large [Pg.173]

An intermediate stage in the purification of human gamma interferon from tissue culture using cation-exchange is shown in Fig. [Pg.174]

A Mono-S stationary phase equilibrated in 10 mM sodium phosphate, pH 7.0 was used with 20% ethylene glycol in the equilibrating buffer. Elution was achieved with a linear salt concentration as shown. [Pg.174]

Insulin Detemir (insulin with a fatty acid molecule attached Chapter 11) [Pg.54]

Increases product serum half-life, thereby reducing the frequency of dosage [Pg.54]

Generation of product that is specifically targeted for uptake by macrophages [Pg.54]

purification of target enzyme to homogeneity and analysis of the amino acid sequence  [Pg.281]

determination and isolation of the corresponding gene, and recombinant expression of protein (steps 1 and 2 are often interchanged)  [Pg.281]

solution of the 3D structure for activity studies, elucidation of the enzyme s mechanism  [Pg.281]

picking of target amino adds for improvement through site-directed mutagenesis  [Pg.281]

site-directed point mutations on the DNA level, doning, and recombinant expression of the mutated gene  [Pg.281]


Because this problem is complex several avenues of attack have been devised in the last fifteen years. A combination of experimental developments (protein engineering, advances in x-ray and nuclear magnetic resonance (NMR), various time-resolved spectroscopies, single molecule manipulation methods) and theoretical approaches (use of statistical mechanics, different computational strategies, use of simple models) [5, 6 and 7] has led to a greater understanding of how polypeptide chains reach the native confonnation. [Pg.2642]

Van Aalten, D.M.F., Findlay, J.B.C., Amadei, A., Berendsen,H.J.C. Essential dynamics of the cellular retinol-binding protein. Evidence for ligand-induced conformational changes. Protein Engin. 8 (1995) 1129-1136. [Pg.35]

Finkelslain A V 1997. Can Protein Unfolding Simulate Protein Folding Protein Engineering 10 843... [Pg.575]

Orengo C A, T P Flores W R Taylor and J M Thornton 1993. Identification and Oassificalion of Prc Fold Families. Protein Engineering 6 485-500. [Pg.577]

Aqvist J, C Medina and J-E Samuelsson 1994. A New Method for Predicting Binding Affinity Computer-aided Drug Design. Protein Engineering 7 385-391. [Pg.649]

Hansson T and J Aqvist 1995. Estimation of Binding Free Energies for HIV Proteinase Inhibitors b Molecular Dynamics Simulations. Protein Engineering 8 1137-1144. [Pg.651]

Biological catalysts — enzymes — are usually proteins. The development of new protein syntheses is nowadays dominated by genetic protein engineering (see section 4.1.2.6). Bio-organic approaches towards novel catalytically active structures and replicating systems try to manage without biopolymers. [Pg.346]

Robson, B. Gamier, J. 1986, Introduction to Proteins and Protein Engineering, Elsevier Amsterdam... [Pg.377]


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