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Instruments / Technical Area	Publication Details	Published	Actions
I04-Macromolecular Crystallography	A monomeric StayGold fluorescent protein Esther Ivorra-Molla , Dipayan Akhuli , Martin B. L. Mcandrew , William Scott , Lokesh Kumar , Saravanan Palani , Masanori Mishima , Allister Crow , Mohan K. Balasubramanian Nature Biotechnology 40 DOI: 10.1038/s41587-023-02018-w Open Access Show Abstract ▼ Abstract: StayGold is an exceptionally bright and stable fluorescent protein that is highly resistant to photobleaching. Despite favorable fluorescence properties, use of StayGold as a fluorescent tag is limited because it forms a natural dimer. Here we report the 1.6 Å structure of StayGold and generate a derivative, mStayGold, that retains the brightness and photostability of the original protein while being fully monomeric.	Dec 2023
I03-Macromolecular Crystallography	Asgard archaea shed light on the evolutionary origins of the eukaryotic ubiquitin-ESCRT machinery Tomoyuki Hatano , Saravanan Palani , Dimitra Papatziamou , Ralf Salzer , Diorge P. Souza , Daniel Tamarit , Mehul Makwana , Antonia Potter , Alexandra Haig , Wenjue Xu , David Townsend , David Rochester , Dom Bellini , Hamdi M. A. Hussain , Thijs J. G. Ettema , Jan Lowe , Buzz Baum , Nicholas P. Robinson , Mohan Balasubramanian Nature Communications 13 DOI: 10.1038/s41467-022-30656-2 Diamond Proposal Number(s): [21426] Open Access Show Abstract ▼ Abstract: The ESCRT machinery, comprising of multiple proteins and subcomplexes, is crucial for membrane remodelling in eukaryotic cells, in processes that include ubiquitin-mediated multivesicular body formation, membrane repair, cytokinetic abscission, and virus exit from host cells. This ESCRT system appears to have simpler, ancient origins, since many archaeal species possess homologues of ESCRT-III and Vps4, the components that execute the final membrane scission reaction, where they have been shown to play roles in cytokinesis, extracellular vesicle formation and viral egress. Remarkably, metagenome assemblies of Asgard archaea, the closest known living relatives of eukaryotes, were recently shown to encode homologues of the entire cascade involved in ubiquitin-mediated membrane remodelling, including ubiquitin itself, components of the ESCRT-I and ESCRT-II subcomplexes, and ESCRT-III and Vps4. Here, we explore the phylogeny, structure, and biochemistry of Asgard homologues of the ESCRT machinery and the associated ubiquitylation system. We provide evidence for the ESCRT-I and ESCRT-II subcomplexes being involved in ubiquitin-directed recruitment of ESCRT-III, as it is in eukaryotes. Taken together, our analyses suggest a pre-eukaryotic origin for the ubiquitin-coupled ESCRT system and a likely path of ESCRT evolution via a series of gene duplication and diversification events.	Jun 2022

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I04-Macromolecular Crystallography

A monomeric StayGold fluorescent protein

Esther Ivorra-Molla , Dipayan Akhuli , Martin B. L. Mcandrew , William Scott , Lokesh Kumar , Saravanan Palani , Masanori Mishima , Allister Crow , Mohan K. Balasubramanian

Nature Biotechnology 40

DOI: 10.1038/s41587-023-02018-w