Gozalbo D, Dubón F, Sentandreu R
Unitat de Microbiología, Facultat de Farmàcia, Universitat de València, Spain.
FEMS Microbiol Lett. 1992 Oct 15;76(3):255-9. doi: 10.1016/0378-1097(92)90345-o.
Subcellular distribution of chitin synthetase has been studied in germ tubes of Candida albicans. Two fractions with synthetase activity were separated from cell homogenates: (i) a mixed membrane fraction where the enzyme, partly in an active form, is associated with the plasma membrane (isopycnic centrifugation of mixed membrane fraction on linear sucrose gradients resolved a unique peak of activity matching with [3H]ConA-labelled membranes at a buoyant density of 1.195 g/ml); and (ii) a cytoplasmic fraction containing fully zymogenic enzyme associated with particles whose buoyant density (determined by isopycnic centrifugation on linear sucrose gradients) depended on the cell breakage conditions. The actual cytoplasmic fraction-enzyme may correspond to particles with buoyant density 1.135 g/ml (chitosomes), whereas the enzyme particles with other densities (1.085 and 1.165 g/ml) probably originated during cell disruption, as has been reported previously to occur during the preparation of yeast cell homogenates.
已对白假丝酵母芽管中的几丁质合成酶的亚细胞分布进行了研究。从细胞匀浆中分离出了两个具有合成酶活性的组分:(i)一个混合膜组分,其中部分呈活性形式的酶与质膜相关(混合膜组分在线性蔗糖梯度上进行等密度离心,分离出一个独特的活性峰,其与浮力密度为1.195 g/ml的[3H]伴刀豆球蛋白A标记的膜相匹配);(ii)一个细胞质组分,含有与颗粒相关的完全无活性的酶原,这些颗粒的浮力密度(通过在线性蔗糖梯度上进行等密度离心测定)取决于细胞破碎条件。实际的细胞质组分 - 酶可能对应于浮力密度为1.135 g/ml的颗粒(几丁质体),而其他密度(1.085和1.165 g/ml)的酶颗粒可能是在细胞破碎过程中产生的,正如先前报道在酵母细胞匀浆制备过程中所发生的那样。