TY  - JOUR
A1  - Makbul, Cihan
A1  - Kraft, Christian
A1  - Grießmann, Matthias
A1  - Rasmussen, Tim
A1  - Katzenberger, Kilian
A1  - Lappe, Melina
A1  - Pfarr, Paul
A1  - Stoffer, Cato
A1  - Stöhr, Mara
A1  - Wandinger, Anna-Maria
A1  - Böttcher, Bettina
T1  - Binding of a pocket factor to Hepatitis B virus capsids changes the rotamer conformation of Phenylalanine 97
JF  - Viruses
N2  - (1) Background: During maturation of the Hepatitis B virus, a viral polymerase inside the capsid transcribes a pre-genomic RNA into a partly double stranded DNA-genome. This is followed by envelopment with surface proteins inserted into a membrane. Envelopment is hypothetically regulated by a structural signal that reports the maturation state of the genome. NMR data suggest that such a signal can be mimicked by the binding of the detergent Triton X 100 to hydrophobic pockets in the capsid spikes. (2) Methods: We have used electron cryo-microscopy and image processing to elucidate the structural changes that are concomitant with the binding of Triton X 100. (3) Results: Our maps show that Triton X 100 binds with its hydrophobic head group inside the pocket. The hydrophilic tail delineates the outside of the spike and is coordinated via Lys-96. The binding of Triton X 100 changes the rotamer conformation of Phe-97 in helix 4, which enables a π-stacking interaction with Trp-62 in helix 3. Similar changes occur in mutants with low secretion phenotypes (P5T and L60V) and in a mutant with a pre-mature secretion phenotype (F97L). (4) Conclusion: Binding of Triton X 100 is unlikely to mimic structural maturation because mutants with different secretion phenotypes show similar structural responses.
KW  - Hepatitis B Virus
KW  - pocket factor
KW  - Triton X 100
KW  - envelopment
KW  - maturation signal
KW  - single strand blocking
KW  - electron cryo-microscopy
KW  - isothermal titration calorimetry
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-248565
SN  - 1999-4915
VL  - 13
IS  - 11
ER  - 
TY  - JOUR
A1  - Laine, Romain F.
A1  - Albecka, Anna
A1  - van de Linde, Sebastian
A1  - Rees, Eric J.
A1  - Crump, Colin M.
A1  - Kaminski, Clemens F.
T1  - Structural analysis of herpes simplex virus by optical super-resolution imaging
JF  - Nature Communications
N2  - Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.
KW  - tegument protein pUL36
KW  - fluorescence microscopy
KW  - monoclonal antibodies
KW  - 3-dimensional structure
KW  - type-1
KW  - nuclear pore complex
KW  - reconstruction microscopy
KW  - localization microscopy
KW  - resolution
KW  - envelopment
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144623
VL  - 6
IS  - 5980
ER  -