Iijima Naoto, Yoshino Hiroto, Ten Lim Chee, Ando Akikazu, Watanabe Katsunori, Nagata Yoshiho
Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo, Chiba 271-8510, Japan.
Biosci Biotechnol Biochem. 2002 Oct;66(10):2083-9. doi: 10.1271/bbb.66.2083.
Our previous studies on the fruit body lectin of Pleurotus cornucopiae revealed the existence of three isolectins, composed of two homodimers and one heterodimer of 16- and 15-kDa subunits. In this study, two genes encoding the lectins were cloned and characterized. Both genes encoded 144 amino acids and only 5 amino acids were different within the coding region, but the nucleotide sequences of the 5'-upstream and 3'-downstream regions differed extensively. Southern hybridization with gene-specific probes showed that one gene encoded the 16-kDa and the other encoded the 15-kDa subunit. Functional lectins were synthesized in Escherichia coli under the direction of these genes. On SDS-PAGE, the recombinant lectins showed the same banding patterns as the native lectins. In amino acid sequence, these lectins showed extensive similarity with the lectin from a nematode-trapping ascomycete fungus, Arthrobotrys oligospora, suggesting that the lectins might also function in capturing nematodes.
我们之前对平菇子实体凝集素的研究表明,存在三种同工凝集素,由两个同型二聚体和一个由16 kDa和15 kDa亚基组成的异型二聚体构成。在本研究中,克隆并鉴定了两个编码凝集素的基因。这两个基因均编码144个氨基酸,编码区内仅有5个氨基酸不同,但5'上游和3'下游区域的核苷酸序列差异很大。用基因特异性探针进行的Southern杂交表明,一个基因编码16 kDa亚基,另一个基因编码15 kDa亚基。在这些基因的指导下,功能性凝集素在大肠杆菌中合成。在SDS-PAGE上,重组凝集素显示出与天然凝集素相同的条带模式。在氨基酸序列上,这些凝集素与一种捕食线虫的子囊菌纲真菌少孢节丛孢菌的凝集素具有广泛的相似性,这表明这些凝集素可能也在捕获线虫中发挥作用。