华中科技大学课件上16 chapter6b.pptVIP

LEHNINGER PRINCIPLES OF BIOCHEMISTRY Fourth Edition Chapter 6 The 3D Structure of Proteins 6.1 Protein Secondary Structure 6.2 Protein Tertiary Structure and Quaternary Structure 6.3 Protein Denaturation and Folding 6.4 Protein-Ligand Interactions 6.5 Working with Proteins Protein Denaturation A native protein conformation is only marginally stable; modest changes in the protein’s environment can bring about structural changes that can affect function. Denaturation (变性)— A loss of three-dimensional structure sufficient to cause loss of function. The denatured state does not necessarily equate with complete unfolding of the protein and randomization of conformation. Under most conditions, denatured proteins exist in a set of partially folded states. What Causes a Protein to be Denatured? Heat— affects the weak interactions in a protein (primarily hydrogen bonds). no covalent bonds are broken If the temperature is increased slowly, a protein’s conformation generally remains intact until an abrupt loss of structure (and function) occurs over a narrow temperature range. Melting temperature (Tm): the midpoint of the temperature range over which denaturation occurs. What Causes a Protein to be Denatured? Extremes of pH alter the net charge on the protein, causing electrostatic repulsion and the disruption of some hydrogen bonding. Organic solvents such as alcohol or acetone, solutes such as urea and guanidine hydrochloride, or by detergents. primarily disrupt the hydrophobic interactions that make up the stable core of globular proteins. Denaturation of Some Proteins is Reversible Denaturation of Some Proteins is Reversible (Cont’d) The refolding of ribonuclease is so accurate that the four intrachain disulfide bonds are re-formed in the same positions in the renatured molecule as in the native ribonuclease. the eight Cys residues could recombine at random to form up to four disulfide bonds in 105 different ways. Anfinsens Dogma Polypeptides Fold Rapidly