Baroja-Fernández E, Zandueta-Criado A, Rodríguez-López M, Akazawa T, Pozueta-Romero J
Instituto de Agrobiotecnologia y Recursos Naturales, Universidad Pública de Navarra/Consejo Superior Investigaciones Cientificas, Mutilva Baja, Spain.
FEBS Lett. 2000 Sep 1;480(2-3):277-82. doi: 10.1016/s0014-5793(00)01951-7.
The intracellular localizations of ADPglucose pyrophosphatase (AGPPase) and ADPglucose pyrophosphorylase (AGPase) have been studied using protoplasts prepared from suspension-cultured cells of sycamore (Acer pseudoplatanus L.). Subcellular fractionation studies revealed that all the AGPPase present in the protoplasts is associated with amyloplasts, whereas more than 60% of AGPase is in the extraplastidial compartment. Immunoblots of amyloplast- and extraplastid-enriched extracts further confirmed that AGPase is located mainly outside the amyloplast. Experiments carried out to identify possible different isoforms of AGPPase in the amyloplast revealed the presence of soluble and starch granule-bound isoforms. We thus propose that ADPglucose levels linked to starch biosynthesis in sycamore cells are controlled by enzymatic reactions catalyzing the synthesis and breakdown of ADPglucose, which take place both inside and outside the amyloplast.
利用从悬铃木(Acer pseudoplatanus L.)悬浮培养细胞制备的原生质体,对ADP葡萄糖焦磷酸酶(AGPPase)和ADP葡萄糖焦磷酸化酶(AGPase)的细胞内定位进行了研究。亚细胞分级分离研究表明,原生质体中存在的所有AGPPase都与造粉体相关,而超过60%的AGPase存在于质体外区室。富含造粉体和质体外提取物的免疫印迹进一步证实,AGPase主要位于造粉体之外。为鉴定造粉体中AGPPase可能存在的不同同工型而进行的实验表明,存在可溶性和淀粉颗粒结合的同工型。因此,我们提出,悬铃木细胞中与淀粉生物合成相关的ADP葡萄糖水平受催化ADP葡萄糖合成和分解的酶促反应控制,这些反应发生在造粉体内外。