Molecular basis of the specific subcellular localization of the C2-like domain of 5-lipoxygenase

Shilpa Kulkarni, Sudipto Das, Colin D. Funk, Diana Murray, Wonhwa Cho

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)13167-13174
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number15
DOIs
StatePublished - Apr 12 2002
Externally publishedYes

Fingerprint

Arachidonate 5-Lipoxygenase
Nuclear Envelope
Membranes
Phosphatidylcholines
Leukotriene A4
Leukotrienes
C2 Domains
Static Electricity
Arachidonic Acid
Tryptophan
Electrostatics
Fluorescence
Chemical activation
Cells
Calcium

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)
  • Molecular Biology
  • Cell Biology

Cite this

Molecular basis of the specific subcellular localization of the C2-like domain of 5-lipoxygenase. / Kulkarni, Shilpa; Das, Sudipto; Funk, Colin D.; Murray, Diana; Cho, Wonhwa.

In: Journal of Biological Chemistry, Vol. 277, No. 15, 12.04.2002, p. 13167-13174.

Research output: Contribution to journalArticle

Kulkarni, Shilpa ; Das, Sudipto ; Funk, Colin D. ; Murray, Diana ; Cho, Wonhwa. / Molecular basis of the specific subcellular localization of the C2-like domain of 5-lipoxygenase. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 15. pp. 13167-13174.
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