Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins
Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithm...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-01-01
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| Series: | Current Research in Structural Biology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665928X22000290 |
| _version_ | 1811292019441532928 |
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| author | Koen Wentinck Christos Gogou Dimphna H. Meijer |
| author_facet | Koen Wentinck Christos Gogou Dimphna H. Meijer |
| author_sort | Koen Wentinck |
| collection | DOAJ |
| description | Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins <100 kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome. |
| first_indexed | 2024-04-13T04:39:06Z |
| format | Article |
| id | doaj.art-cbe1c58cf7a1480084ede770ded8d289 |
| institution | Directory Open Access Journal |
| issn | 2665-928X |
| language | English |
| last_indexed | 2024-04-13T04:39:06Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Current Research in Structural Biology |
| spelling | doaj.art-cbe1c58cf7a1480084ede770ded8d2892022-12-22T03:02:05ZengElsevierCurrent Research in Structural Biology2665-928X2022-01-014332337Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteinsKoen Wentinck0Christos Gogou1Dimphna H. Meijer2Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, van der Maasweg 9, 2629HZ, Delft, the NetherlandsDepartment of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, van der Maasweg 9, 2629HZ, Delft, the NetherlandsCorresponding author.; Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, van der Maasweg 9, 2629HZ, Delft, the NetherlandsSignificant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins <100 kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome.http://www.sciencedirect.com/science/article/pii/S2665928X22000290 |
| spellingShingle | Koen Wentinck Christos Gogou Dimphna H. Meijer Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins Current Research in Structural Biology |
| title | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
| title_full | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
| title_fullStr | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
| title_full_unstemmed | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
| title_short | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
| title_sort | putting on molecular weight enabling cryo em structure determination of sub 100 kda proteins |
| url | http://www.sciencedirect.com/science/article/pii/S2665928X22000290 |
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