Enzyme 2课件.pptVIP

7. Factors Affecting Enzyme Activity The catalytic activity of enzymes is influenced by numerous factors: --substrate concentration --enzyme concentration --temperature, pH -- inhibitors --activators (1) Effect of [E] on velocity Optimum pH (2) Effect of pH on E activity pH can affect the ionization of E、S and ES. There is a optimum pH for each E, at which E activity is maximum. pH 5～8 is optimum for most enzyme (3) Effect of temperature on E activity (3) Effect of temperature on E activity most of E : optimum T around 25~40℃ T＜ optimum T: increase in the v with increasing T T ＞ optimum T: increasing T can decrease the v, the enzyme is going to be denatured Inhibitors decrease E activity, but not denature the E protein Reversible inhibitors (可逆性抑制剂) Irreversible inhibitors (不可逆性抑制剂) Reversible inhibitor (可逆性抑制剂): inhibitor non-covalently binds to E protein and decreases the E activity. --competitive inhibition (竞争性抑制作用) --noncompetitive inhibition (非竞争性抑制作用) --uncompetitive inhibition (反竞争性抑制作用) You should know: The features of inhibitors The changes of Km and Vmax in the presence of inhibitors The Linerweave-Burk plot of inhibition the inhibitor(I) structure is similar to S The I reversibly competitively binds to the active site of E (1) Competitive inhibition** I only binds to free E and competes with S Vmax: unchanged apparent Km (Kmapp,表观Km): increased affinity of E for S is decreased Inhibition is reversible as high[S] competes for I 竞争性抑制在临床上的应用 细菌: 喋呤定 + 对氨基苯甲酸 + 谷氨酸 二氢叶酸 四氢叶酸 二氢叶酸还原酶 人可以直接从食物中得到叶酸 竞争性抑制在临床的应用 抗癌药物 氨甲喋呤（MIX)---叶酸 -----竞争性抑制二氢叶酸还原酶 5-氟尿嘧啶（5-FU)---胸嘧啶(T) 6-巯基嘌呤（6-MP)---鸟嘌呤(A) ----竞争性抑制相应核苷酸代谢酶---抑制DNA和RNA的生物合成 细胞死亡 the I binds reversibly at a site other than the active site pure noncompetitive inhibition** The I binds at a different site to the S Vmax: decreased apparent Km (Kmap