Sychrová H, Horák J, Kotyk A
Department of Membrane Transport, Czechoslovak Academy of Sciences, Prague.
Biochim Biophys Acta. 1989 Jan 30;978(2):203-8. doi: 10.1016/0005-2736(89)90116-8.
Systems of L-lysine transport in Schizosaccharomyces pombe are not constitutive, as at no phase of growth in a rich medium is lysine taken up. Transport activity appears only after preincubation of harvested cells with glucose or another suitable source of energy. If cycloheximide is added during this preincubation no transport systems are synthesized. After removal of glucose, the activity of the transport system decays with a half-time of 13 min. The transport of L-lysine into S. pombe cells from the stationary phase of growth preincubated for 60 min with 1% D-glucose is mediated by at least two systems, the high-affinity one with a Kt of 26 mumol/l and Jmax of 4.95 nmol/min per mg dry wt., the low-affinity one with a KT of 1.1 mmol/l and Jmax of 11.8 nmol/min per mg dry wt. The transport of lysine mediated by these two systems proceeds uphill. The high-affinity system has a pH optimum at 4.0-4.2, the accumulation ratio is highest at a cell density 2-5 mg dry wt. per ml and decreases with increasing lysine concentrations. Lysine accumulated by this system does not exit from cells. The only potent competitive inhibitors are L-arginine, L-histidine and D-lysine. The other amino acids tested do not behave as competitive inhibitors. Of the various metabolic inhibitors tested, the most potent were proton conductors and antimycin A.
粟酒裂殖酵母中的L-赖氨酸转运系统并非组成型的,因为在丰富培养基中生长的任何阶段,赖氨酸都不会被吸收。只有在收获的细胞用葡萄糖或其他合适的能量来源进行预孵育后,转运活性才会出现。如果在该预孵育过程中加入环己酰亚胺,则不会合成转运系统。去除葡萄糖后,转运系统的活性以13分钟的半衰期衰减。从用1% D-葡萄糖预孵育60分钟的生长稳定期的细胞中,L-赖氨酸向粟酒裂殖酵母细胞内的转运至少由两个系统介导,高亲和力系统的Kt为26 μmol/l,Jmax为每毫克干重4.95 nmol/分钟,低亲和力系统的KT为1.1 mmol/l,Jmax为每毫克干重11.8 nmol/分钟。由这两个系统介导的赖氨酸转运是逆浓度梯度进行的。高亲和力系统的最适pH为4.0 - 4.2,在细胞密度为每毫升2 - 5毫克干重时积累率最高,并随赖氨酸浓度增加而降低。通过该系统积累的赖氨酸不会从细胞中流出。唯一有效的竞争性抑制剂是L-精氨酸、L-组氨酸和D-赖氨酸。所测试的其他氨基酸不表现为竞争性抑制剂。在测试的各种代谢抑制剂中,最有效的是质子导体和抗霉素A。