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Hijacking prenyl and geranyl transferases - A route to carry out click modifications and to enhance cellular permeability of peptides

劫持异戊二烯基和香叶基转移酶 - 进行点击修饰并增强肽的细胞渗透性的途径

基本信息

批准号：
EP/S027246/1
负责人：
Wael Houssen
金额：
$ 131.57万
依托单位：
University of Aberdeen
依托单位国家：
英国
项目类别：
Fellowship
财政年份：
2019
资助国家：
英国
起止时间：
2019 至无数据
项目状态：
未结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FS027246%2F1
关键词：
Hijacking prenyl geranyl transferases route

项目摘要

Recent advances in biological research have allowed a better understanding of the causation of many diseases and identified new targets for therapy. An ideal drug should bind to a specific cellular target and have no affinity to others. One of the recently identified challenging targets for drug discovery is the protein-protein interactions that have been proved to be involved in many difficult-to-treat diseases e.g. immune disorders and cancer. These interactions are taking place along the extended surface of large proteins and thus are very challenging for small molecule drugs. Biological drugs e.g. antibodies are large molecules and can disrupt protein-protein interactions but cannot be administered orally and are very expensive. Macrocyclic peptides are an emerging class of drug candidates that have the ability to disrupt protein-protein interactions e.g. the immune-suppressant, cyclosporin that made transplant surgery possible. They are smaller in size and are very much cheaper than biologics. In contrast to their linear "non-cyclic" counterparts, they are more stable against enzymes and are semi-rigid to fit better with their targets much like a key fits into a lock. A limitation that hampers the development of many of these compounds is their low ability to cross cellular membranes and to reach intracellular targets. Several modified cyclic peptides are commonly found in medicinal natural products. These compounds were evolved via natural selection which is presumably driven by their pharmacological potency against specific molecular targets as well as their ability to reach these targets that is, by crossing one or more biological membranes. In nature, several modifications are introduced to cyclic peptides to enhance membrane permeability. These modifications aim to reduce the hydrophilic (polar) surface of the molecule by shielding with hydrophobic side chains thus the compound can easily diffuse through the hydrophobic (mainly lipid) cellular membranes. Ideally these modifications should be applied to specific sites to avoid a large reduction in water solubility or the change of three dimensional shape of the molecule with subsequent decrease in its ability to bind to its target. Recent research revealed how a large group of these modified cyclic peptides is made inside their hosts. In this project, I will identify and recruit new modifying biosynthetic enzymes that will add hydrophobic chemical groups such as prenyl and geranyl groups at specific sites in cyclic peptides. Making these modifications using chemical methods is very challenging, not eco-friendly and in most cases entails total re-synthesis which is time consuming. I will determine the structure and biochemical features of these enzymes to identify the key residues that underlie their activity and specificity. I will use these insights to engineer and generate enzyme variants with different residue specificity and ability to introduce other chemical groups. I will use chemical synthesis and the engineered enzymes to generate modified derivatives of bespoke bioactive cyclic peptides and test the effect of different modifications on membrane permeability and the three dimensional shape of the molecule that underlies target affinity. These data will help to generate a computational model to predict membrane permeability of bioactive cyclic peptides that will be invaluable for development of peptides into drugs.

生物学研究的最新进展使人们能够更好地了解许多疾病的病因，并确定了新的治疗目标。理想的药物应该结合到特定的细胞靶点，而与其他药物没有亲和力。最近发现的具有挑战性的药物发现目标之一是蛋白质-蛋白质相互作用，已被证明与许多难以治疗的疾病有关，如免疫疾病和癌症。这些相互作用发生在大蛋白质的延伸表面上，因此对小分子药物来说是非常具有挑战性的。生物药物，如抗体，是大分子，可以破坏蛋白质之间的相互作用，但不能口服，而且非常昂贵。大环肽是一类新兴的候选药物，具有干扰蛋白质-蛋白质相互作用的能力，例如使移植手术成为可能的免疫抑制剂环孢素。它们的体积更小，而且比生物制品便宜得多。与线性的“非循环”同行相比，它们对酶更稳定，并且是半刚性的，以更好地适应他们的目标，就像钥匙放入锁中一样。阻碍这些化合物发展的一个限制是它们穿过细胞膜和到达细胞内靶点的能力很低。在药用天然产物中常见几种修饰的环肽。这些化合物是通过自然选择进化而来的，这可能是由于它们对特定分子靶标的药理效力以及它们达到这些靶标的能力，即通过穿过一个或多个生物膜。在自然界中，环肽被引入几种修饰来增强膜的通透性。这些修饰旨在通过疏水侧链的屏蔽来减少分子的亲水(极性)表面，从而使化合物能够很容易地通过疏水(主要是脂质)细胞膜扩散。理想情况下，这些修饰应该应用于特定的位置，以避免水溶解度的大幅降低或分子三维形状的改变，从而降低其与目标的结合能力。最近的研究揭示了这些修饰的环肽是如何在它们的宿主体内产生的。在这个项目中，我将鉴定和招募新的修饰生物合成酶，这些酶将在环肽的特定位置添加疏水化学基团，如异戊烯和香叶基。使用化学方法进行这些修饰非常具有挑战性，不环保，而且在大多数情况下需要完全重新合成，这是非常耗时的。我将确定这些酶的结构和生化特征，以确定支撑其活性和特异性的关键残基。我将利用这些洞察力来设计和产生具有不同残基特异性和引入其他化学基团的能力的酶变体。我将使用化学合成和工程酶来产生定制的生物活性环肽的修饰衍生物，并测试不同修饰对膜通透性和作为靶标亲和力基础的分子的三维形状的影响。这些数据将有助于建立一个计算模型来预测活性环肽的膜透过性，这将对多肽的药物开发具有非常重要的价值。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Biochemical characterization of a cyanobactin arginine-N-prenylase from the autumnalamide biosynthetic pathway.

DOI：
10.1039/d2cc01799g
发表时间：
2022-10-27
期刊：
CHEMICAL COMMUNICATIONS
影响因子：
4.9
作者：
Clemente, Claudia;Johnson, Nicholas;Ouyang, Xiaodan;Popin, Rafael, V;Dall'Angelo, Sergio;Wahlsten, Matti;Jokela, Jouni;Colombano, Alessandro;Nardone, Brunello;Fewer, David P.;Houssen, Wael E.
通讯作者：
Houssen, Wael E.

Exploring the Limits of Cyanobactin Macrocyclase PatGmac: Cyclization of PawS-Derived Peptide Sunflower Trypsin Inhibitor-1 and Cyclotide Kalata B1.

DOI：
10.1021/acs.jnatprod.2c01158
发表时间：
2023-03-24
期刊：
JOURNAL OF NATURAL PRODUCTS
影响因子：
5.1
作者：
Muhammad, Taj;Houssen, Wael E.;Thomas, Louise;Alexandru-Crivac, Cristina-Nicoleta;Gunasekera, Sunithi;Jaspars, Marcel;Goransson, Ulf
通讯作者：
Goransson, Ulf

Chemoenzymatic Late-Stage Modifications Enable Downstream Click-Mediated Fluorescent Tagging of Peptides

化学酶后期修饰实现下游点击介导的肽荧光标记