H.-H. Jang et al. / Archives of Biochemistry and Biophysics 493 (2010) 143–150
149
0
.5 mol%inplasmaandsubcellularmembranes.PAisalsoinvolvedin
(D
2–4)CYP1B1 than the (
D
2–26) enzyme. This result may be as-
diversecellularfunctionssuchascellgrowth,proliferation,reproduc-
tion,andresponsestohormones[33].
cribed to a deletion of Trp residue in N-terminal anchor region of
2–26) enzyme. We could not also exclude the possibility that
(D
Interestingly, the results for PA-mediated functional regulations
of CYP1B1 parallel the previous observations that PA increased the
catalytic activities of CYP1A2 and CYP3A4 with increasing the lipid
concentrations in membranes [29,30]. It was also demonstrated
that CYP2B1 induced clustering of the lipid PA to a greater extent
than other anionic phospholipids did, and that PA increased the
ability of CYP2B1 to interact with lipid monolayers [34]. Based
on these observations, we anticipate that PA and/or PA-induced
membrane properties play an important role in the interaction be-
tween various P450 enzymes and membranes. In addition, the
requirement of anionic phospholipids for the efficient function of
P450 enzymes was suggested previously [20]. At present, it is rel-
evant that CYP1B1 interacts with PA and the biological functions
can be regulated by the phospholipid. It should be also considered
that the concentrations PA used in the present study seem to be
higher than those found in cells. However, as we suggested previ-
ously, the mixing property of PA is not random but the phase sep-
aration occurs in the liquid–crystalline phase of membranes [35]
through intermolecular hydrogen bonding [36]. In addition, PA
concentration is very fluctuant as a signaling lipid depending on
cellular states.
other Trp residues in CYP1B1 exert their influence on the energy
transfer and thus the results as well as conformational change re-
vealed by Trp fluorescence (Fig. 4C and D) might not provide spe-
cific understanding of how the protein is interacting with the
membrane. Nonetheless, the energy transfer result suggests that
CYP1B1 has the ability to bind and penetrate pre-formed mem-
brane bilayers depending on the type of phospholipids present in
the membrane.
Acknowledgments
This work was supported in part by the 21C Frontier Microbial
Genomics and the Application Center Program of the Ministry of
Education, Science and Technology of the Republic of Korea; the
Korea Science and Engineering Foundation (Grant R01-2008-000-
2
1072-02008); and the Second Stage BK21 Project from the Minis-
try of Education, Science and Technology of the Republic of Korea.
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