PMID- 26906727
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20230720
IS  - 1432-1017 (Electronic)
IS  - 0175-7571 (Print)
IS  - 0175-7571 (Linking)
VI  - 45
IP  - 4
DP  - 2016 May
TI  - Measuring kinetic drivers of pneumolysin pore structure.
PG  - 365-76
LID - 10.1007/s00249-015-1106-x [doi]
AB  - Most membrane attack complex-perforin/cholesterol-dependent cytolysin (MACPF/CDC) 
      proteins are thought to form pores in target membranes by assembling into 
      pre-pore oligomers before undergoing a pre-pore to pore transition. Assembly 
      during pore formation is into both full rings of subunits and incomplete rings 
      (arcs). The balance between arcs and full rings is determined by a mechanism 
      dependent on protein concentration in which arc pores arise due to kinetic 
      trapping of the pre-pore forms by the depletion of free protein subunits during 
      oligomerization. Here we describe the use of a kinetic assay to study pore 
      formation in red blood cells by the MACPF/CDC pneumolysin from Streptococcus 
      pneumoniae. We show that cell lysis displays two kinds of dependence on protein 
      concentration. At lower concentrations, it is dependent on the pre-pore to pore 
      transition of arc oligomers, which we show to be a cooperative process. At higher 
      concentrations, it is dependent on the amount of pneumolysin bound to the 
      membrane and reflects the affinity of the protein for its receptor, cholesterol. 
      A lag occurs before cell lysis begins; this is dependent on oligomerization of 
      pneumolysin. Kinetic dissection of cell lysis by pneumolysin demonstrates the 
      capacity of MACPF/CDCs to generate pore-forming oligomeric structures of variable 
      size with, most likely, different functional roles in biology.
FAU - Gilbert, Robert J C
AU  - Gilbert RJ
AD  - Division of Structural Biology, Wellcome Trust Centre for Human Genetics, 
      University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK. 
      gilbert@strubi.ox.ac.uk.
FAU - Sonnen, Andreas F-P
AU  - Sonnen AF
AD  - European Molecular Biology Laboratory, Structural and Computational Biology Unit, 
      Meyerhofstrasse 1, 69117, Heidelberg, Germany.
LA  - eng
GR  - WT_/Wellcome Trust/United Kingdom
GR  - 090532/Z/09/Z/WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160223
PL  - Germany
TA  - Eur Biophys J
JT  - European biophysics journal : EBJ
JID - 8409413
RN  - 0 (Bacterial Proteins)
RN  - 0 (Streptolysins)
RN  - 0 (plY protein, Streptococcus pneumoniae)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/chemistry/metabolism
MH  - Erythrocytes/metabolism
MH  - Kinetics
MH  - Porosity
MH  - Sheep
MH  - Streptolysins/*chemistry/*metabolism
PMC - PMC4823331
OTO - NOTNLM
OT  - Kinetics
OT  - MACPF/CDC
OT  - Membrane structure
OT  - Oligomerization
OT  - Pore formation
OT  - Toroidal pore
EDAT- 2016/02/26 06:00
MHDA- 2016/12/15 06:00
PMCR- 2016/02/23
CRDT- 2016/02/25 06:00
PHST- 2015/10/07 00:00 [received]
PHST- 2015/12/07 00:00 [accepted]
PHST- 2015/12/01 00:00 [revised]
PHST- 2016/02/25 06:00 [entrez]
PHST- 2016/02/26 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
PHST- 2016/02/23 00:00 [pmc-release]
AID - 10.1007/s00249-015-1106-x [pii]
AID - 1106 [pii]
AID - 10.1007/s00249-015-1106-x [doi]
PST - ppublish
SO  - Eur Biophys J. 2016 May;45(4):365-76. doi: 10.1007/s00249-015-1106-x. Epub 2016 
      Feb 23.