Pub Date : 2023-07-07DOI: 10.1107/s2053273323096420
Stefan A. Kolek, Patrick Shaw Stewart, Jack Stubbs, P. Baldock
Serial data collection usually requires relatively small crystals that are well-ordered. Microseeding is an effective way to generate such samples. During the ten years since the random microseed matrix-screening (rMMS) method was published, understanding of the theoretical advantages of the method has increased [2 4], and several practical variations of the method have emerged. Moreover seeding can be carried out in a microbatch-underoil setup, which is easy to scale up, volume-wise, and allows easy interpretation of phase diagrams. By combining these techniques, control can be increased and sample quality for both routine and advanced data collection improved. Protein structure determination by cryoEM requires expensive equipment that has low throughput. It is therefore wasteful to examine samples that can be shown in advance to be aggregated, since such samples are unlikely to be suitable. We used a high-throughput screening approach with dynamic light scattering to explore 96 chemical conditions with as little as 10 μL of protein solution to identify conditions with reduced aggregation.
{"title":"Sample preparation for routine and advanced structural biology, including serial data collection, MicroED and cryo-EM","authors":"Stefan A. Kolek, Patrick Shaw Stewart, Jack Stubbs, P. Baldock","doi":"10.1107/s2053273323096420","DOIUrl":"https://doi.org/10.1107/s2053273323096420","url":null,"abstract":"Serial data collection usually requires relatively small crystals that are well-ordered. Microseeding is an effective way to generate such samples. During the ten years since the random microseed matrix-screening (rMMS) method was published, understanding of the theoretical advantages of the method has increased [2 4], and several practical variations of the method have emerged. Moreover seeding can be carried out in a microbatch-underoil setup, which is easy to scale up, volume-wise, and allows easy interpretation of phase diagrams. By combining these techniques, control can be increased and sample quality for both routine and advanced data collection improved. Protein structure determination by cryoEM requires expensive equipment that has low throughput. It is therefore wasteful to examine samples that can be shown in advance to be aggregated, since such samples are unlikely to be suitable. We used a high-throughput screening approach with dynamic light scattering to explore 96 chemical conditions with as little as 10 μL of protein solution to identify conditions with reduced aggregation.","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"198 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323099023
Jieun Kim
{"title":"Crystal structure of human interleukin-2 in complex with TCB2, a new antibody-drug candidate with antitumor activity","authors":"Jieun Kim","doi":"10.1107/s2053273323099023","DOIUrl":"https://doi.org/10.1107/s2053273323099023","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"198 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323098534
Vinu V. Panikkattu, Viraj De Silvia, Christer Aakeroy
{"title":"From molecular to supramolecular to functional materials","authors":"Vinu V. Panikkattu, Viraj De Silvia, Christer Aakeroy","doi":"10.1107/s2053273323098534","DOIUrl":"https://doi.org/10.1107/s2053273323098534","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323096183
Vanessa Bijak, Michael Gucwa, Michal Szcygiel, K. Handing, Wladek Minor
{"title":"Analysis of human serum albumin metal complexes","authors":"Vanessa Bijak, Michael Gucwa, Michal Szcygiel, K. Handing, Wladek Minor","doi":"10.1107/s2053273323096183","DOIUrl":"https://doi.org/10.1107/s2053273323096183","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"26 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323098078
D. Tchoń, Aleksandra Zwolenik, Anna M. Makal
Diamond anvil cells (DACs) exert high pressure while allowing light to access the sample. They enable single crystals to be investigated under extreme conditions by means of X - ray diffraction (XRD). Structure factors collected this way provide unique structural information, but their quality is never on par with routine experiments.1,2 A typical 35° opening angle of modern DACs renders up to 97% of the limiting sphere inaccessible. Resulting diffraction patterns are thus systematically incomplete, especially in samples with low internal symmetry, which impedes the crystal structure solution and affects the applicability of techniques that require high reciprocal space coverage.3,4
金刚石砧座（DAC）在施加高压的同时允许光线进入样品。通过这些装置，可以在极端条件下利用 X 射线衍射（XRD）对单晶体进行研究。通过这种方法收集的结构因子可提供独特的结构信息，但其质量永远无法与常规实验相提并论1,2。因此，得出的衍射图样往往是不完整的，尤其是在内部对称性较低的样品中，这阻碍了晶体结构的求解，并影响了需要高倒易空间覆盖率的技术的适用性。
{"title":"Increasing completeness in single-crystal high-pressure diffraction experiments by pre-orienting crystals","authors":"D. Tchoń, Aleksandra Zwolenik, Anna M. Makal","doi":"10.1107/s2053273323098078","DOIUrl":"https://doi.org/10.1107/s2053273323098078","url":null,"abstract":"Diamond anvil cells (DACs) exert high pressure while allowing light to access the sample. They enable single crystals to be investigated under extreme conditions by means of X - ray diffraction (XRD). Structure factors collected this way provide unique structural information, but their quality is never on par with routine experiments.1,2 A typical 35° opening angle of modern DACs renders up to 97% of the limiting sphere inaccessible. Resulting diffraction patterns are thus systematically incomplete, especially in samples with low internal symmetry, which impedes the crystal structure solution and affects the applicability of techniques that require high reciprocal space coverage.3,4","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"13 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361509","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323098388
M. T. Swulius
{"title":"Imaging neurons by cryo-electron tomography","authors":"M. T. Swulius","doi":"10.1107/s2053273323098388","DOIUrl":"https://doi.org/10.1107/s2053273323098388","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"24 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323097991
Hong Zhou
The last decade has witnessed advances in high-resolution cryoEM “revolutionizing” structural biology; as such, cryoEM has become a highly sought-after means of biochemical and biomedical investigations. Results from cryoEM are beginning to significantly enhance our understanding of the cellular processes responsible for maintenance, transmission and expression of genetic information at the atomic level. At the heart of these processes lie macromolecular complexes, within and outside the cell, which can now be studied by cryoEM. Understanding how and why these complexes function relies on visualizing their three-dimensional (3D) structures at their endogenous and multiple functional states. Towards this end, we have developed an integrative proteomics cryoEM methods to determine atomic structures of native cellular complexes, sub - particle refinement and nucleic acid modeling methods to model genomic RNA and DNA in action. Of particular note, our cryoID method (Ho et al., Nat Methods, 2020) allows determination of atomic structures of endogenous complexes in cellular milieu, capturing their multiple states, including those in their acts of carrying out their functions. Applications of cryoID has enabled atomic structure determination of previously intractable biological systems from cellular milieu and membrane.
{"title":"New cryo-EM methods to capture endogenous complexes in multiple functional states at atomic resolution","authors":"Hong Zhou","doi":"10.1107/s2053273323097991","DOIUrl":"https://doi.org/10.1107/s2053273323097991","url":null,"abstract":"The last decade has witnessed advances in high-resolution cryoEM “revolutionizing” structural biology; as such, cryoEM has become a highly sought-after means of biochemical and biomedical investigations. Results from cryoEM are beginning to significantly enhance our understanding of the cellular processes responsible for maintenance, transmission and expression of genetic information at the atomic level. At the heart of these processes lie macromolecular complexes, within and outside the cell, which can now be studied by cryoEM. Understanding how and why these complexes function relies on visualizing their three-dimensional (3D) structures at their endogenous and multiple functional states. Towards this end, we have developed an integrative proteomics cryoEM methods to determine atomic structures of native cellular complexes, sub - particle refinement and nucleic acid modeling methods to model genomic RNA and DNA in action. Of particular note, our cryoID method (Ho et al., Nat Methods, 2020) allows determination of atomic structures of endogenous complexes in cellular milieu, capturing their multiple states, including those in their acts of carrying out their functions. Applications of cryoID has enabled atomic structure determination of previously intractable biological systems from cellular milieu and membrane.","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"42 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323098789
Emily Armbruster
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-
{"title":"Transitioning from academia to a cryo-EM CRO","authors":"Emily Armbruster","doi":"10.1107/s2053273323098789","DOIUrl":"https://doi.org/10.1107/s2053273323098789","url":null,"abstract":"-","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"44 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323097656
Tony Hu
{"title":"Hydrogen-bonded frameworks for molecular structure determination","authors":"Tony Hu","doi":"10.1107/s2053273323097656","DOIUrl":"https://doi.org/10.1107/s2053273323097656","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"41 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-07DOI: 10.1107/s2053273323097255
Ashley Weiland
{"title":"Optimizing data collection time for absolute configuration determination","authors":"Ashley Weiland","doi":"10.1107/s2053273323097255","DOIUrl":"https://doi.org/10.1107/s2053273323097255","url":null,"abstract":"","PeriodicalId":6903,"journal":{"name":"Acta Crystallographica Section A Foundations and Advances","volume":"35 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139361637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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