Rosa J C, De Oliveira P S, Garratt R, Beltramini L, Resing K, Roque-Barreira M C, Greene L J
Centro de Química de Proteínas and Departamento de Ginecologia e Obstetricia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Brazil.
Protein Sci. 1999 Jan;8(1):13-24. doi: 10.1110/ps.8.1.13.
The complete amino acid sequence of the lectin KM+ from Artocarpus integrifolia (jackfruit), which contains 149 residues/mol, is reported and compared to those of other members of the Moraceae family, particularly that of jacalin, also from jackfruit, with which it shares 52% sequence identity. KM+ presents an acetyl-blocked N-terminus and is not posttranslationally modified by proteolytic cleavage as is the case for jacalin. Rather, it possesses a short, glycine-rich linker that unites the regions homologous to the alpha- and beta-chains of jacalin. The results of homology modeling implicate the linker sequence in sterically impeding rotation of the side chain of Asp141 within the binding site pocket. As a consequence, the aspartic acid is locked into a conformation adequate only for the recognition of equatorial hydroxyl groups on the C4 epimeric center (alpha-D-mannose, alpha-D-glucose, and their derivatives). In contrast, the internal cleavage of the jacalin chain permits free rotation of the homologous aspartic acid, rendering it capable of accepting hydrogen bonds from both possible hydroxyl configurations on C4. We suggest that, together with direct recognition of epimeric hydroxyls and the steric exclusion of disfavored ligands, conformational restriction of the lectin should be considered to be a new mechanism by which selectivity may be built into carbohydrate binding sites. Jacalin and KM+ adopt the beta-prism fold already observed in two unrelated protein families. Despite presenting little or no sequence similarity, an analysis of the beta-prism reveals a canonical feature repeatedly present in all such structures, which is based on six largely hydrophobic residues within a beta-hairpin containing two classic-type beta-bulges. We suggest the term beta-prism motif to describe this feature.
报道了波罗蜜(木菠萝)凝集素KM +的完整氨基酸序列,其含有149个残基/摩尔,并与桑科家族的其他成员进行了比较,特别是同样来自木菠萝的jacalin,与之具有52%的序列同一性。KM +呈现乙酰化封闭的N末端,并且不像jacalin那样通过蛋白水解切割进行翻译后修饰。相反,它具有一个短的、富含甘氨酸的连接子,该连接子将与jacalin的α链和β链同源的区域连接起来。同源性建模的结果表明,连接子序列在空间上阻碍了结合位点口袋内Asp141侧链的旋转。因此,天冬氨酸被锁定在仅适合识别C4差向异构中心上的赤道羟基(α-D-甘露糖、α-D-葡萄糖及其衍生物)的构象中。相比之下,jacalin链的内部切割允许同源天冬氨酸自由旋转,使其能够接受来自C4上两种可能的羟基构型的氢键。我们认为,连同对差向异构羟基的直接识别和对不利配体的空间排斥一起,凝集素的构象限制应被视为一种可将选择性构建到碳水化合物结合位点的新机制。Jacalin和KM +采用了在两个不相关的蛋白质家族中已经观察到的β-棱柱折叠。尽管几乎没有序列相似性,但对β-棱柱的分析揭示了所有此类结构中反复出现的一个典型特征,该特征基于包含两个经典型β-凸起的β-发夹内的六个主要疏水残基。我们建议用术语β-棱柱基序来描述这个特征。