Department of Microbiology, University of Illinois, Urbana, Illinois, USA.
Department of Biochemistry, University of Illinois, Urbana, Illinois, USA.
Proteins. 2024 Apr;92(4):435-448. doi: 10.1002/prot.26642. Epub 2023 Nov 23.
Biotin (vitamin H or B7) is a coenzyme essential for all forms of life. Biotin has biological activity only when covalently attached to a few key metabolic enzyme proteins. Most organisms have only one attachment enzyme, biotin protein ligase (BPL), which attaches biotin to all target proteins. The sequences of these proteins and their substrate proteins are strongly conserved throughout biology. Structures of both the biotin ligase- and biotin-acceptor domains of mammals, plants, several bacterial species, and archaea have been determined. These, together with mutational analyses of ligases and their protein substrates, illustrate the exceptional specificity of this protein modification. For example, the Escherichia coli BPL biotinylates only one of the >4000 cellular proteins. Several bifunctional bacterial biotin ligases transcriptionally regulate biotin synthesis and/or transport in concert with biotinylation. The human BPL has been demonstrated to play an important role in that mutations in the BPL encoding gene cause one form of the disease, biotin-responsive multiple carboxylase deficiency. Promiscuous mutant versions of several BPL enzymes release biotinoyl-AMP, the active intermediate of the ligase reaction, to solvent. The released biotinoyl-AMP acts as a chemical biotinylation reagent that modifies lysine residues of neighboring proteins in vivo. This proximity-dependent biotinylation (called BioID) approach has been heavily utilized in cell biology.
生物素(维生素 H 或 B7)是所有生命形式必需的辅酶。生物素只有与少数关键代谢酶蛋白共价结合时才具有生物活性。大多数生物体只有一种附着酶,生物素蛋白连接酶(BPL),它将生物素附着到所有靶蛋白上。这些蛋白质及其底物蛋白质的序列在整个生物学中都强烈保守。哺乳动物、植物、几种细菌物种和古细菌的生物素连接酶和生物素受体结构域的结构已被确定。这些结构,以及对连接酶及其蛋白质底物的突变分析,说明了这种蛋白质修饰的非凡特异性。例如,大肠杆菌 BPL 仅生物素化 >4000 种细胞蛋白中的一种。几种双功能细菌生物素连接酶与生物素化协同转录调节生物素合成和/或运输。已经证明人类 BPL 在该疾病的一种形式中起重要作用,即生物素反应性多羧化酶缺乏症。几种 BPL 酶的混杂突变体将生物素酰基-AMP(连接酶反应的活性中间体)释放到溶剂中。释放的生物素酰基-AMP 作为一种化学生物素化试剂,在体内修饰邻近蛋白质的赖氨酸残基。这种依赖于接近的生物素化(称为 BioID)方法在细胞生物学中得到了广泛应用。