Crystals in Minutes: Instant On‐Site Microcrystallisation of Various Flavours of the CYP102A1 (P450BM3) Haem Domain

Stanfield Joshua Kyle; Omura Keita; Matsumoto Ayaka; Kasai Chie; Sugimoto Hiroshi; Shiro Yoshitsugu; Watanabe Yoshihito; Shoji Osami

摘要

Abstract > Despite CYP102A1 (P450BM3) representing one of the most extensively researched metalloenzymes, crystallisation of its haem domain upon modification can be a challenge. Crystal structures are indispensable for the efficient structure‐based design of P450BM3 as a biocatalyst. The abietane diterpenoid derivative N‐abietoyl‐ <sc>l</sc> ‐tryptophan (AbiATrp) is an outstanding crystallisation accelerator for the wild‐type P450BM3 haem domain, with visible crystals forming within 2?hours and diffracting to a near‐atomic resolution of 1.22??. Using these crystals as seeds in a cross‐microseeding approach, an assortment of P450BM3 haem domain crystal structures, containing previously uncrystallisable decoy molecules and diverse artificial metalloporphyrins binding various ligand molecules, as well as heavily tagged haem‐domain variants, could be determined. Some of the structures reported herein could be used as models of different stages of the P450BM3 catalytic cycle. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> > 尽管CYP102A1（P450BM3）代表了最广泛研究的金属酶之一，但在改性后其血液结构域的结晶可能是一个挑战。晶体结构对于P450BM3的高效结构设计是一种生物催化剂是必不可少的。双替代萜类衍生物n-abietoyl- <sc> l </ sc> -Tryptophan（AbiaTrp）是野生型P450bm3血液结构域的出色结晶促进剂，在2℃下形成可见晶体，并衍射到近似原子分辨率为1.22。使用这些晶体作为交叉微缩合方法的种子，可以确定含有先前未易于算可的诱饵分子和不同的人造金属卟啉的各种各样的P450BM3血管结构域晶体结构，以及重叠各种配体分子以及重标记的血液结构域变体。本文报道的一些结构可以用作P450BM3催化循环的不同阶段的模型。 </ p> </摘要> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-19011/'>《Angewandte Chemie》</a> <b style="margin: 0 2px;">|</b><span>2020年第19期</span><b style="margin: 0 2px;">|</b><span>共8页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Stanfield Joshua Kyle&option=202" target="_blank" rel="nofollow">Stanfield Joshua Kyle;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Omura Keita&option=202" target="_blank" rel="nofollow">Omura Keita;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Matsumoto Ayaka&option=202" target="_blank" rel="nofollow">Matsumoto Ayaka;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Kasai Chie&option=202" target="_blank" rel="nofollow">Kasai Chie;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sugimoto Hiroshi&option=202" target="_blank" rel="nofollow">Sugimoto Hiroshi;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shiro Yoshitsugu&option=202" target="_blank" rel="nofollow">Shiro Yoshitsugu;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Watanabe Yoshihito&option=202" target="_blank" rel="nofollow">Watanabe Yoshihito;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shoji Osami&option=202" target="_blank" rel="nofollow">Shoji Osami;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Department of ChemistryNagoya University Furo-cho Chikusa-kuNagoya 464-8602 Japan;</p> <p>Department of ChemistryNagoya University Furo-cho Chikusa-kuNagoya 464-8602 Japan;</p> <p>Department of ChemistryNagoya University Furo-cho Chikusa-kuNagoya 464-8602 Japan;</p> <p>Department of ChemistryNagoya University Furo-cho Chikusa-kuNagoya 464-8602 Japan;</p> <p>RIKEN SPring-8 Centre1-1-1 Kouto Sayo Hyogo 679-5148 Japan;</p> <p>Graduate School of Life ScienceUniversity of Hyogo3-2-1-Kouto Kamigori-cho Ako-gun Hyogo 678-1297 Japan;</p> <p>Research Center for Materials ScienceNagoya University Furo-cho Chikusa-kuNagoya 464-860 Japan;</p> <p>Department of ChemistryNagoya University Furo-cho Chikusa-kuNagoya 464-8602 Japan;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/1188.html" title="应用化学">应用化学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=crystal growth&option=203" rel="nofollow">crystal growth;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=cytochromes&option=203" rel="nofollow">cytochromes;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=decoy molecules&option=203" rel="nofollow">decoy molecules;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=enzyme models&option=203" rel="nofollow">enzyme models;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=protein structures&option=203" rel="nofollow">protein structures;</a> </p> <div class="translation"> 机译：晶体生长;细胞学;诱饵分子;酶模型;蛋白质结构; </div> </li> </ul> </div> </div> <div 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