Bahbouhi Bouchaib, Chazal Nathalie, Seidah Nabil Georges, Chiva Cristina, Kogan Marcelo, Albericio Fernando, Giralt Ernest, Bahraoui Elmostafa
Laboratoire d'Immuno-Virologie, EA 30-38 Université Paul Sabatier, UFR/SVT, 118 route de Narbonne 31062 Toulouse, France.
Biochem J. 2002 Sep 15;366(Pt 3):863-72. doi: 10.1042/BJ20020052.
The aim of the present study was to evaluate the capacity of synthetic l- and d-peptides encompassing the HIV-1(BRU) gp160 REKR cleavage site to interfere with HIV and simian immuno-deficiency virus (SIV) replication and maturation of the envelope glycoprotein (Env) precursors. To facilitate their penetration into cells, a decanoyl (dec) group was added at the N-terminus. The sequences synthesized included dec5d or dec5l (decREKRV), dec9d or dec9l (decRVVQREKRV) and dec14d or dec14l (TKAKRRVVQREKRV). The peptide dec14d was also prepared with a chloromethane (cmk) group as C-terminus. Because l-peptides exhibit significant cytotoxicity starting at 35 microM, further characterization was conducted mostly with d-peptides, which exhibited no cytotoxicity at concentrations higher than 70 microM. The data show that only dec14d and dec14dcmk could inhibit HIV-1(BRU), HIV-2(ROD) and SIV(mac251) replication and their syncytium-inducing capacities. Whereas peptides dec5d and dec9d were inactive, dec14dcmk was at least twice as active as peptide dec14d. At the molecular level, our data show a direct correlation between anti-viral activity and the ability of the peptides to interfere with maturation of the Env precursors. Furthermore, we show that when tested in vitro the dec14d peptide inhibited PC7 with an inhibition constant K(i)=4.6 microM, whereas the peptide dec14l preferentially inhibited furin with a K(i)=28 microM. The fact that PC7 and furin are the major prohormone convertases reported to be expressed in T4 lymphocytes, the principal cell targets of HIV, suggests that they are involved in the maturation of HIV and SIV Env precursors.
本研究的目的是评估包含HIV-1(BRU) gp160 REKR裂解位点的合成L型和D型肽干扰HIV和猿猴免疫缺陷病毒(SIV)复制以及包膜糖蛋白(Env)前体成熟的能力。为便于其进入细胞,在N端添加了一个癸酰基(dec)基团。合成的序列包括dec5d或dec5l(decREKRV)、dec9d或dec9l(decRVVQREKRV)以及dec14d或dec14l(TKAKRRVVQREKRV)。肽dec14d还制备了氯甲烷(cmk)基团作为C端。由于L型肽在35微摩尔时就表现出显著的细胞毒性,因此进一步的表征主要使用D型肽进行,D型肽在浓度高于70微摩尔时未表现出细胞毒性。数据表明,只有dec14d和dec14dcmk能够抑制HIV-1(BRU)、HIV-2(ROD)和SIV(mac251)的复制及其诱导合胞体的能力。而肽dec5d和dec9d无活性,dec14dcmk的活性至少是肽dec14d的两倍。在分子水平上,我们的数据表明抗病毒活性与肽干扰Env前体成熟的能力之间存在直接相关性。此外,我们表明,在体外测试时,dec14d肽抑制PC7的抑制常数K(i)=4.6微摩尔,而肽dec14l优先抑制弗林蛋白酶,K(i)=28微摩尔。PC7和弗林蛋白酶是据报道在HIV的主要细胞靶标T4淋巴细胞中表达的主要前激素转化酶,这一事实表明它们参与了HIV和SIV Env前体的成熟。