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SUMMARY:Contr. talk  - Production and purification of the K+-dependent P-T
 ype ATPase KdpFABC for crystallization and cryo-EM
DTSTART;VALUE=DATE-TIME:20210525T141000Z
DTEND;VALUE=DATE-TIME:20210525T143000Z
DTSTAMP;VALUE=DATE-TIME:20260721T193202Z
UID:indico-contribution-184-1135@lindico453.srv.lu.se
DESCRIPTION:Speakers: Charlott Stock (Aarhus University)\nAt very low extr
 acellular K+ concentrations (<100 μM)\, KdpFABC rescues bacterial cells b
 y taking up K+ against an up to 105-fold concentration gradient. To unders
 tand the mechanism of this heterotetrameric complex\, KdpFABC was\npurifie
 d for functional and structural investigations. A new expression system fo
 r KdpFABC was generated\, a 3-step chromatographic purification implemente
 d and protein functionality in different detergents as well as in the pres
 ence of\ninhibitors was tested. The work was performed in the lab of Prof.
  Inga Hänelt at the University of Frankfurt from 2014-2020.\n\nhttps://li
 ndico453.srv.lu.se/event/219/contributions/1135/
LOCATION:
URL:https://lindico453.srv.lu.se/event/219/contributions/1135/
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SUMMARY:Keynote talk - Preparation of lipid flippases for cryo-EM studies
DTSTART;VALUE=DATE-TIME:20210525T133000Z
DTEND;VALUE=DATE-TIME:20210525T141000Z
DTSTAMP;VALUE=DATE-TIME:20260721T193202Z
UID:indico-contribution-184-1134@lindico453.srv.lu.se
DESCRIPTION:Speakers: Joseph Lyons (Aarhus University\, Denmark)\nP4-ATPas
 es are lipid flippases that drive the ATP-dependent inward translocation (
 flipping) of lipids within the membrane. They are members of the P-type AT
 Pase superfamily and largely function as binary complexes with an auxiliar
 y protein from the CDC50 family. Select lipid flippases are auto-regulated
  by conserved motifs in their termini and require activation by an externa
 l stimulus\, i.e.\, binding of an activating lipid or regulatory protein\,
  and/or specific phosphorylation of the termini.  To elucidate the auto-re
 gulation and transport mechanisms\, we determined the structures of yeast 
 and mammalian lipid flippases by cryo-electron microscopy. \n\nIn this tal
 k\, I will give an overview of the tour de force we embarked on\, from exp
 ression through purification to cryo-EM imaging of these lipid flippases.\
 n\nhttps://lindico453.srv.lu.se/event/219/contributions/1134/
LOCATION:
URL:https://lindico453.srv.lu.se/event/219/contributions/1134/
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