几丁质酶及其研究进展

本文从几丁质酶的分布、发育调节、可诱导性、分子生物学及抗病基因工程等方面近年来的进展进行了综合论述,并对其进一步的应用提出展望。     

几丁质酶的研究进展

几丁质是自然界中含量仅次于纤维素的第二大类多糖,已从各种细菌、真菌、植物、哺乳动物和昆虫中分离并鉴定出来。随着近年来对几丁质及其衍生物的了解,发现此类多糖的物理化学特性在现实中有非常广泛的应用。几丁质酶是一类分解几丁质的酶,广泛分布于细菌、真菌、植物、哺乳动物、昆虫等物种中,并发挥着重要作用。近年来国内外工作者对这些不同来源的几丁质酶做了深入研究,为其在医学和农业上更广泛的应用提供理论基础。     

微生物几丁质酶研究进展

微生物几丁质酶不仅在生物降解几丁质方面起着重要作用,而且可通过水解病原真菌的细胞壁而有效地抑制其生长。到目前为止,人们已经分离和克隆出了大量的微生物几丁质酶及其基因。尽管这些几丁质酶各不相同,但它们却具有类同的蛋白质结构域:信号肽、催化结构域和几丁质结合结构域等。本文着重介绍几丁质酶的结构和分子特征、表达和调控机理,并且分析了该酶的应用前景。     

细菌几丁质酶研究进展

真菌病害是影响作物产量的重要原因 ,而几丁质酶能有效抑制其生长、水解其细胞壁。对研究较多的细菌几丁质酶及几丁质酶基因的分子生物学研究进展进行了综述 ,并对细菌几丁质酶基因利用存在的问题进行了探讨。     

酸性哺乳动物几丁质酶的研究进展

酸性哺乳动物几丁质酶是食用燕窝中普遍且大量存在的活性蛋白质之一,是哺乳动物中常见的几丁质酶,能够在酸性环境下保持活性并水解几丁质。酸性哺乳动物几丁质酶主要在人体的呼吸系统和消化系统中表达,且在多种病理机制和免疫炎症中有调节作用。本文综述了酸性哺乳动物几丁质酶的理化性质与功能,包括其在人体各个系统和器官中发挥的多种作用及相关机制,以探究其对人类健康和疾病的影响。     

昆虫几丁质酶的研究进展

综述了几丁质酶的特性,生理作用,基因结构和表达及其在植物保护和基因工程中的应用。     

植物几丁质酶的研究进展

几丁质酶是植物抗真菌基因工程的热点之一。本文叙述了植物几丁质酶的特性、结构和功能、基因结构;按最新资料对以前的植物几丁质酶的分类系统进行了完善;概述了几丁质酶的分子进化的各家观点及其模型,并归纳了植物几丁质酶的生物学作用。     

苏云金芽孢杆菌几丁质酶的研究进展

苏云金芽孢杆菌制剂作为无公害农药已经得到社会的认可,如果再开发其几丁质酶抑制真菌和杀虫增效功能,不仅可充分利用这一农业微生物菌种资源,也将给予传统生物农药以新的生命力。综述了苏云金芽孢杆菌几丁质酶方面研究的国内外最新进展。     

植物几丁质酶的研究进展

自 1 92 1年Ｆｏｌｐｍｅｒｓ首次从细菌和放线菌中证实水溶性几丁质酶存在以来 ,研究者又相继在多种微生物、植物、动物中分离到几丁质酶。在高等植物中 ,几丁质酶分布于草本植物和木本植物或单子叶植物和双子叶植物 ,主要存在于植物的根、茎、叶、花、果实、种子及胚等部位 ,种子、花器、根中的几丁质酶含量一般高于其它器官[1] 。现以在玉米、水稻、小麦、大麦、棉花、油菜、甜橙等 70多种栽培植物和野生植物中检测到几丁质酶活性[2 ] 。1　几丁质酶的特性及分类定位1 1　几丁质酶的一般特性几丁质酶是一种糖苷酶 ,以几丁质 [(1 ,4) - 2…     

Acetazolamide-based fungal chitinase inhibitors

Chitin is an essential structural component of the fungal cell wall. Chitinases are thought to be important for fungal cell wall remodelling, and inhibition of these enzymes has been proposed as a potential strategy for development of novel anti-fungals. The fungal pathogen Aspergillus fumigatus possesses two distinct multi-gene chitinase families. Here we explore acetazolamide as a chemical scaffold for the inhibition of an A. fumigatus 'plant-type' chitinase. A co-crystal structure of AfChiA1 with acetazolamide was used to guide synthesis and screening of acetazolamide analogues that yielded SAR in agreement with these structural data. Although acetazolamide and its analogues are weak inhibitors of the enzyme, they have a high ligand efficiency and as such are interesting leads for future inhibitor development.     

Phage display screening for peptidic chitinase inhibitors

A phage display library with disulfide-cyclized peptides was screened for peptides binding to chitinases from Serratia marcescens. One of those peptides was found to efficiently inhibit chitinase A and two others were inhibitors of chitinase B. Complete substitutional analysis of all three peptides using cellulose-bound peptide spot synthesis revealed key interaction positions and allowed optimization of the chitinase B inhibitory peptides towards higher affinity, with inhibitory constants in the lower nanomolar range. Inhibition by all peptides proved to be competitive and highly specific for the chitinase used to select them, as shown with a series of chitinases from different organisms.     

Screening-based discovery of Aspergillus fumigatus plant-type chitinase inhibitors

A limited therapeutic arsenal against increasing clinical disease due to Aspergillus spp. necessitates urgent characterisation of new antifungal targets. Here we describe the discovery of novel, low micromolar chemical inhibitors of Aspergillus fumigatus family 18 plant-type chitinase A1 (AfChiA1) by high-throughput screening (HTS). Analysis of the binding mode by X-ray crystallography confirmed competitive inhibition and kinetic studies revealed two compounds with selectivity towards fungal plant-type chitinases. These inhibitors provide new chemical tools to probe the effects of chitinase inhibition on A. fumigatus growth and virulence, presenting attractive starting points for the development of further potent drug-like molecules.     

Recent advances on cyclins, CDKs and CDK inhibitors

In eukaryotes, cell division is controlled by cyclin-dependent kinases (CDKs). Here we summarize a few new developments on the regulation of the cell cycle by CDK-cyclin complexes. We have focused on three aspects in which there has been recent progress: the structural analysis of these complexes, the phenotypes of mice carrying knockouts of CDK inhibitors and the role of proteolysis in the regulation of the cell cycle.     

肉毒神经毒素抑制剂的研究进展

肉毒神经毒素(botulinum neurotoxins,BoNTs)是由梭状芽孢杆菌属分泌的外毒素,是目前已知毒性最强的生物类毒素.BoNTs共分为7种血清型(A-G),其中A型导致的肉毒中毒最为常见.由于肉毒毒素的强毒性及易于制备,其已被列为A类生物恐怖制剂.目前,针对肉毒中毒的有效治疗手段为早期注射抗毒素血清.但...     

Structure-based virtual screening of highly potent inhibitors of the nematode chitinase CeCht1

Nematode chitinases play vital roles in various physiological processes, including egg hatching, larva moulting, and reproduction. Small-molecule inhibitors of nematode chitinases have potential applications for controlling nematode pests. On the basis of the crystal structure of CeCht1, a representative chitinase indispensable to the eggshell chitin degradation of the model nematode Caenorhabditis elegans, we have discovered a series of novel inhibitors bearing a (R)-3,4-diphenyl-4,5-dihydropyrrolo[3,4-c]pyrazol-6(2H)-one scaffold by hierarchical virtual screening. The crystal structures of CeCht1 complexed with two of these inhibitors clearly elucidated their interactions with the enzyme active site. Based on the inhibitory mechanism, several analogues with improved inhibitory activities were identified, among which the compound PP28 exhibited the most potent activity with a K_i value of 0.18 μM. This work provides the structural basis for the development of novel nematode chitinase inhibitors.     

LIM域结合蛋白研究进展

近来,几组研究人员分别独立地分离出一种新的蛋白质,先后将其命名为LDB、NLI和CLIM,统称为LIM域结合蛋白,它在爪蟾和小鼠的胚胎以及成体小鼠和人中有着广泛的表达,并能与多种LIM同源域蛋白发生相互作用,进而在神经系统发育及红细胞和淋巴细胞的成熟过程中发挥作用,LIM域结合蛋白在果蝇中的同源性CHIP/dLDB的发现,为进一步研究LIM同源域蛋白提供了有价值的线索。     

Potent family-18 chitinase inhibitors: x-ray structures, affinities, and binding mechanisms

Six novel inhibitors of Vibrio harveyi chitinase A (VhChiA), a family-18 chitinase homolog, were identified by in vitro screening of a library of pharmacologically active compounds. Unlike the previously identified inhibitors that mimicked the reaction intermediates, crystallographic evidence from 14 VhChiA-inhibitor complexes showed that all of the inhibitor molecules occupied the outer part of the substrate-binding cleft at two hydrophobic areas. The interactions at the aglycone location are well defined and tightly associated with Trp-397 and Trp-275, whereas the interactions at the glycone location are patchy, indicating lower affinity and a loose interaction with two consensus residues, Trp-168 and Val-205. When Trp-275 was substituted with glycine (W275G), the binding affinity toward all of the inhibitors dramatically decreased, and in most structures two inhibitor molecules were found to stack against Trp-397 at the aglycone site. Such results indicate that hydrophobic interactions are important for binding of the newly identified inhibitors by the chitinase. X-ray data and isothermal microcalorimetry showed that the inhibitors occupied the active site of VhChiA in three different binding modes, including single-site binding, independent two-site binding, and sequential two-site binding. The inhibitory effect of dequalinium in the low nanomolar range makes this compound an extremely attractive lead compound for plausible development of therapeutics against human diseases involving chitinase-mediated pathologies.