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芹菜叶柄细胞膜的体外葡聚糖合成:膜在决定所形成连接类型中的作用。

In vitro glucan synthesis by membranes of celery petioles: the role of the membrane in determining the type of linkage formed.

作者信息

Jacob S R, Northcote D H

出版信息

J Cell Sci Suppl. 1985;2:1-11. doi: 10.1242/jcs.1985.supplement_2.1.

Abstract

Glucan synthesis was achieved with an in vitro membrane fraction from the petioles of celery (Apium graveolens). The optimum conditions for maximum synthesis were established. The Km and Vmax for the enzymic system were 1.0 mM and 0.19 microM min-1 mg protein-1, respectively. Mechanical damage to the membrane fraction altered the proportion of beta-(1----3) to beta-(1----4) glucosyl linkages that were synthesized. We suggest that cellulose synthesis (beta-(1----4)-linked glucan chains) is controlled by the availability of UDP-glucose at the plasma membrane surface in conjunction with an organized relationship between the synthase system and a specifically oriented glucosyl radical acting as an acceptor held on the membrane surface. An intact membrane is therefore necessary to direct synthesis for the beta-(1----4) bond by an enzyme that is capable of transglucosylation to the secondary alcoholic groups on C-2, C-3 or C-4 of the acceptor radical. The specificity of the system is controlled by the whole enzyme complex held on the membrane.

摘要

利用芹菜(旱芹)叶柄的体外膜部分实现了葡聚糖的合成。确定了最大合成的最佳条件。该酶系统的米氏常数(Km)和最大反应速度(Vmax)分别为1.0 mM和0.19 μmol min-1 mg蛋白质-1。对膜部分的机械损伤改变了所合成的β-(1→3)与β-(1→4)葡糖基连接的比例。我们认为,纤维素合成(β-(1→4)连接的葡聚糖链)受质膜表面尿苷二磷酸葡萄糖(UDP-葡萄糖)可用性的控制,同时受合成酶系统与作为受体固定在膜表面的特定取向葡糖基自由基之间有组织关系的控制。因此,完整的膜对于由能够将转葡糖基作用于受体自由基C-2、C-3或C-4上的仲醇基团的酶来指导β-(1→4)键的合成是必要的。该系统的特异性由固定在膜上的整个酶复合物控制。

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