TY - JOUR
T1 - Molecular basis of the specific subcellular localization of the C2-like domain of 5-lipoxygenase
AU - Kulkarni, Shilpa
AU - Das, Sudipto
AU - Funk, Colin D.
AU - Murray, Diana
AU - Cho, Wonhwa
PY - 2002/4/12
Y1 - 2002/4/12
N2 - The activation of 5-lipoxygenase (5-LO) involves its calcium-dependent translocation to the nuclear envelope, where it catalyzes the two-step transformation of arachidonic acid into leukotriene A4, leading to the synthesis of various leukotrienes. To understand the mechanism by which 5-LO is specifically targeted to the nuclear envelope, we studied the membrane binding properties of the amino-terminal domain of 5-LO, which has been proposed to have a C2 domain-like structure. The model building, electrostatic potential calculation, and in vitro membrane binding studies of the isolated C2-like domain of 5-LO and selected mutants show that this Ca2+-dependent domain selectively binds zwitterionic phosphatidylcholine, which is conferred by tryptophan residues (Trp13, Trp75, and Trp102) located in the putative Ca2+-binding loops. The spatiotemporal dynamics of the enhanced green fluorescence protein-tagged C2-like domain of 5-LO and mutants in living cells also show that the phosphatidylcholine selectivity of the C2-like domain accounts for the specific targeting of 5-LO to the nuclear envelope. Together, these results show that the C2-like domain of 5-LO is a genuine Ca2+-dependent membrane-targeting domain and that the subcellular localization of the domain is governed in large part by its membrane binding properties.
AB - The activation of 5-lipoxygenase (5-LO) involves its calcium-dependent translocation to the nuclear envelope, where it catalyzes the two-step transformation of arachidonic acid into leukotriene A4, leading to the synthesis of various leukotrienes. To understand the mechanism by which 5-LO is specifically targeted to the nuclear envelope, we studied the membrane binding properties of the amino-terminal domain of 5-LO, which has been proposed to have a C2 domain-like structure. The model building, electrostatic potential calculation, and in vitro membrane binding studies of the isolated C2-like domain of 5-LO and selected mutants show that this Ca2+-dependent domain selectively binds zwitterionic phosphatidylcholine, which is conferred by tryptophan residues (Trp13, Trp75, and Trp102) located in the putative Ca2+-binding loops. The spatiotemporal dynamics of the enhanced green fluorescence protein-tagged C2-like domain of 5-LO and mutants in living cells also show that the phosphatidylcholine selectivity of the C2-like domain accounts for the specific targeting of 5-LO to the nuclear envelope. Together, these results show that the C2-like domain of 5-LO is a genuine Ca2+-dependent membrane-targeting domain and that the subcellular localization of the domain is governed in large part by its membrane binding properties.
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U2 - 10.1074/jbc.M112393200
DO - 10.1074/jbc.M112393200
M3 - Article
C2 - 11796736
AN - SCOPUS:0037066761
SN - 0021-9258
VL - 277
SP - 13167
EP - 13174
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 15
ER -