Katahira Riko, Ashihara Hiroshi
Department of Advanced Bioscience, Graduate School of Humanities and Sciences, Ochanomizu University, Tokyo, 112-8610, Japan.
Planta. 2006 Dec;225(1):115-26. doi: 10.1007/s00425-006-0334-9. Epub 2006 Jul 15.
To find general metabolic profiles of purine ribo- and deoxyribonucleotides in potato (Solanum tuberosum L.) plants, we looked at the in situ metabolic fate of various (14)C-labelled precursors in disks from growing potato tubers. The activities of key enzymes in potato tuber extracts were also studied. Of the precursors for the intermediates in de novo purine biosynthesis, [(14)C]formate, [2-(14)C]glycine and [2-(14)C]5-aminoimidazole-4-carboxyamide ribonucleoside were metabolised to purine nucleotides and were incorporated into nucleic acids. The rates of uptake of purine ribo- and deoxyribonucleosides by the disks were in the following order: deoxyadenosine > adenosine > adenine > guanine > guanosine > deoxyguanosine > inosine > hypoxanthine > xanthine > xanthosine. The purine ribonucleosides, adenosine and guanosine, were salvaged exclusively to nucleotides, by adenosine kinase (EC 2.7.1.20) and inosine/guanosine kinase (EC 2.7.1.73) and non-specific nucleoside phosphotransferase (EC 2.7.1.77). Inosine was also salvaged by inosine/guanosine kinase, but to a lesser extent. In contrast, no xanthosine was salvaged. Deoxyadenosine and deoxyguanosine, was efficiently salvaged by deoxyadenosine kinase (EC 2.7.1.76) and deoxyguanosine kinase (EC 2.7.1.113) and/or non-specific nucleoside phosphotransferase (EC 2.7.1.77). Of the purine bases, adenine, guanine and hypoxanthine but not xanthine were salvaged for nucleotide synthesis. Since purine nucleoside phosphorylase (EC 2.4.2.1) activity was not detected, adenine phosphoribosyltransferase (EC 2.4.2.7) and hypoxanthine/guanine phosphoribosyltransferase (EC 2.4.2.8) seem to play the major role in salvage of adenine, guanine and hypoxanthine. Xanthine was catabolised by the oxidative purine degradation pathway via allantoin. Activity of the purine-metabolising enzymes observed in other organisms, such as purine nucleoside phosphorylase (EC 2.4.2.1), xanthine phosphoribosyltransferase (EC 2.4.2.22), adenine deaminase (EC 3.5.4.2), adenosine deaminase (EC 3.5.4.4) and guanine deaminase (EC 3.5.4.3), were not detected in potato tuber extracts. These results suggest that the major catabolic pathways of adenine and guanine nucleotides are AMP --> IMP --> inosine --> hypoxanthine --> xanthine and GMP --> guanosine --> xanthosine --> xanthine pathways, respectively. Catabolites before xanthosine and xanthine can be utilised in salvage pathways for nucleotide biosynthesis.
为了探究马铃薯(Solanum tuberosum L.)植株中嘌呤核糖核苷酸和脱氧核糖核苷酸的一般代谢概况,我们研究了来自生长中的马铃薯块茎切片中各种(14)C标记前体的原位代谢命运。我们还研究了马铃薯块茎提取物中关键酶的活性。在嘌呤从头生物合成中间体的前体中,[(14)C]甲酸、[2-(14)C]甘氨酸和[2-(14)C]5-氨基咪唑-4-甲酰胺核糖核苷被代谢为嘌呤核苷酸并掺入核酸中。切片对嘌呤核糖核苷和脱氧核糖核苷的摄取速率顺序如下:脱氧腺苷>腺苷>腺嘌呤>鸟嘌呤>鸟苷>脱氧鸟苷>肌苷>次黄嘌呤>黄嘌呤>黄苷。嘌呤核糖核苷腺苷和鸟苷仅通过腺苷激酶(EC 2.7.1.20)、肌苷/鸟苷激酶(EC 2.7.1.73)和非特异性核苷磷酸转移酶(EC 2.7.1.77)挽救为核苷酸。肌苷也可被肌苷/鸟苷激酶挽救,但程度较小。相比之下,黄苷未被挽救。脱氧腺苷和脱氧鸟苷通过脱氧腺苷激酶(EC 2.7.1.76)、脱氧鸟苷激酶(EC 2.7.1.113)和/或非特异性核苷磷酸转移酶(EC 2.7.1.77)有效挽救。在嘌呤碱基中,腺嘌呤、鸟嘌呤和次黄嘌呤而非黄嘌呤被挽救用于核苷酸合成。由于未检测到嘌呤核苷磷酸化酶(EC 2.4.2.1)的活性,腺嘌呤磷酸核糖转移酶(EC 2.4.2.7)和次黄嘌呤/鸟嘌呤磷酸核糖转移酶(EC 2.4.2.8)似乎在腺嘌呤、鸟嘌呤和次黄嘌呤的挽救中起主要作用。黄嘌呤通过氧化嘌呤降解途径经尿囊素分解代谢。在马铃薯块茎提取物中未检测到在其他生物体中观察到的嘌呤代谢酶的活性,如嘌呤核苷磷酸化酶(EC 2.4.2.1)、黄嘌呤磷酸核糖转移酶(EC 2.4.2.22)、腺嘌呤脱氨酶(EC 3.5.4.2)、腺苷脱氨酶(EC 3.5.4.4)和鸟嘌呤脱氨酶(EC 3.5.4.3)。这些结果表明,腺嘌呤和鸟嘌呤核苷酸的主要分解代谢途径分别是AMP→IMP→肌苷→次黄嘌呤→黄嘌呤和GMP→鸟苷→黄苷→黄嘌呤途径。黄苷和黄嘌呤之前的分解代谢产物可用于核苷酸生物合成的挽救途径。