H. Decock et al., IN-VITRO INSERTION AND ASSEMBLY OF OUTER-MEMBRANE PROTEIN PHOE OF ESCHERICHIA-COLI K-12 INTO THE OUTER-MEMBRANE - ROLE OF TRITON X-100, The Journal of biological chemistry, 271(22), 1996, pp. 12885-12890
The assembly of the in vitro synthesized outer membrane protein PhoE i
nto purified outer membranes was investigated, The assembly appeared t
o be strongly stimulated by the presence of low amounts of Triton X-10
0 (optimal 0.08%, w/v). The role of Triton X-100 in the in vitro syste
m was further examined. Pretreating outer membranes with Triton X-100
did not make the membranes competent for correct assembly, indicating
that the detergent did not act on the membrane but at the protein leve
l. PhoE became assembly-incompetent with a half-life of approximately
12 min and 90 s at 37 degrees C in the absence and presence, respectiv
ely, of 0.08% Triton X-100, Apparently, Triton X-100 induces an assemb
ly-competent state in the PhoE protein with a very short half-life. Fu
rthermore, the efficiency of correct assembly of PhoE was greatly redu
ced when outer membranes of deep rough lipopolysaccharide mutants were
used, indicating an important role of lipopolysaccharides in the asse
mbly of the porin.