Download Document

IT’S ALIVE!: THE
PHENOMENON OF VIRAL
BREATHING
STRUCTURAL JOURNAL CLUB
AUGUST 25, 2011
WHAT IS VIRAL
BREATHING?
Breathing is when the capsid undergoes reversible
conformational changes not induced by an outside source.
Example: An eternal epitope is exposed on the capsid
surface periodically without the presence
of receptor.
DIGESTION TIME
COURSE
CRYSTAL STRUCTURE
OF VP4
DIGESTION IN THE
PRESENCE OF DRUG
DIGESTION WITH
ANTIBODY
STABILITY OF
DIGESTION
Mutant:
V1188M/
C1199W
INTERNAL
NEUTRALIZATION SITES
VP4
VP1 N
terminus
TEMPERATURE
DEPENDENCE
REVERSIBILITY OF
INTERNAL
NEUTRALIZATION
NEUTRALIZATION BY
VP4 ANTIBODY
CROSS REACTIVITY
OF ANTIBODIES
CVB3 STRAIN CPE
Three strains of CVB3:
-
28 – cause complete
CPE on HeLa but
not RD,
representative strain
-
RD – DAF binding
strain
-
RDVa – variant of
RD which emerged
from carrier culture
HEMAGGLUTINATION
(DAF BINDING)
LIGAND BINDING OF
CVB3
DAF
CAR
CVB3/28
CVB3/RD
CVB3/RDVa
GROWTH CURVES
HeLa
CVB3/28
RD
RD Carrier
CVB3/RD
CVB3/RDVa
CVB3 DECAY
CVB3/28
CVB3/RD
CVB3/RDVa
AMINO ACID
CHANGES IN CVB3
WHAT WE HAVE DONE
Incubated CVB3-RD with CAR at 4 °C, RT, and 37 °C and saw
the following:
WHAT WE WANT TO DO
• Repeat Carson’s decay experiment to determine what the
virus is doing to loose infectivity
• Negstain of different time points
• Gel of time points
• Western of time points
• Eventually would like to put it on affinity grid
• Virus on lipid alone without receptor to determine
whether it will undergo conformation change on lipid
alone
• Receptor doesn’t bind all over the virus – only one
position
NOW FOR SOMETHING
COMPLETELY DIFFERENT!
WARNING: YOU SHOULD NOT TRY THE
FOLLOWING AT HOME! YOU WILL NEVER
BE ABLE TO PUBLISH THIS!
135S AND 135S-P1
80S-P1 COMPLEXES
P1 FAB ON PV
1 OR 8 PARTICLE
RECONSTRUCTION