IBT01 L2 Proteins
IBT01 L2 Proteins
20 standard amino acids At physiological pH amino acids are charged as shown on left Can act either as acid or base Soluble in water
Why?
Peptide Bond
Lys (K)
pKa = 10.5 Below 10.5 pH it will be +vely charged
Asp
(D)3.9 Glu (E)4.3 Arg (R)12.0 Lys (K)10.5 His (H)6.08 (+vely charged below 6.08)
The pH-dependence of the activity displayed by enzymes and the pH-dependence of protein stability, for example, are properties that are determined by the pKa values of amino acid side chains.
Hydrophobic/Nonpolar
Polar
Hydrophobic/Nonpolar
e = molar absorbtivity coefficient (Molar Extinction Coefficient ) with units of L mol-1 cm-1 Means how strongly a substance absorbs light Intrinsic property of substance, depends on chemical nature.
A = no units c = mol L-1 l = cm
Biocatalyst Biochemical reactions Industrial application: Proteases/Lipases = added detergents Cellulase = stone wash jeans
Pharmaceutical Industry
Genetic blood disorder characterized by red blood cells that assume an abnormal, rigid, sickle shape. Mutation in hemoglobin
Human CJD disease: Cruetzfeld-Jakob disease Cattle: Mad cow disease Prion diseases are transmissible from host to host of a single species and, sometimes, even from one species to another (such as a laboratory animal) Destroy brain tissue giving it a spongy appearance
Remember: When purifying a protein one has to be careful that it does not get denatured. Heating = denatures protein SDS (sodium dodecyl sulfate) (detergent) = denatures protein Urea (high concentrations denature proteins
Separation techniques make use of protein properties Properties of protein depend on the amino acid sequence
(pI)
The net charge (the algebraic sum of all the charged groups present) of any amino acid, peptide or protein, will depend upon the pH of the surrounding aqueous environment. As the pH of a solution of an amino acid or protein changes so too does the net charge. When the net charge of an amino acid or protein is zero the pH will be equivalent to the isoelectric point: pI.
Histidine
+
Below pH 6
Above pH 6
Protonated
H+
~6.0 pKa
OH-
Unprotonated
Below pH4.1
Above pH4.1
Protonated
H+
pKa
OH-
Unprotonated
+
Protonated
H+ H+
12.5
pKa
OHOH-
0
Unprotonated
The peptide bond is not charged, however you still have a + at the N-terminal, - at the Cterminal, and any charges on side chains An amino acid can not exist as COOH/NH2 because any pH low enough to protonate the COO- group would also protonate the NH2
9.9
2.3
4.3 6
12.4
+vely charged
pI
deprotonation [OH-]
pH above pI
-vely charge