ATP-binding Cassette Transporter A1 Contains an NH2-terminal Signal Anchor Sequence That Translocates the Protein's First Hydrophilic Domain to the Exoplasmic Space

Michael L. Fitzgerald, Armando J Mendez, Kathryn J. Moore, Lorna P. Andersson, Hess A. Panjeton, Mason W. Freeman

Research output: Contribution to journalArticle

102 Citations (Scopus)

Abstract

Mutations in the ATP-binding cassette transporter A1 (ABCA1) transporter are associated with Tangier disease and a defect in cellular cholesterol efflux. The amino terminus of the ABCA1 transporter has two putative in-frame translation initiation sites, 60 amino acids apart. A cluster of hydrophobic amino acids form a potentially cleavable signal sequence in this 60-residue extension. We investigated the functional role of this extension and found that it was required for stable protein expression of transporter constructs containing any downstream transmembrane domains. The extension directed transporter translocation across the ER membrane with an orientation that resulted in glycosylation of amino acids immediately distal to the signal sequence. Neither the native signal sequence nor a green fluorescent protein tag, fused at the amino terminus, was cleaved from ABCA1. The green fluorescent protein fusion protein had efflux activity comparable with wild type ABCA1 and demonstrated a predominantly plasma membrane distribution in transfected cells. These data establish a requirement for the upstream 60 amino acids of ABCA1. This region contains an uncleaved signal anchor sequence that positions the amino terminus in a type II orientation leading to the extracellular presentation of an ∼600-amino acid loop in which loss-of-function mutations cluster in Tangier disease patients.

Original languageEnglish
Pages (from-to)15137-15145
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number18
StatePublished - May 4 2001

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ATP-Binding Cassette Transporters
Protein Sorting Signals
Anchors
Amino Acids
Tangier Disease
Green Fluorescent Proteins
Proteins
Glycosylation
Mutation
Cell membranes
Fusion reactions
Cholesterol
Cell Membrane
Membranes
Defects

ASJC Scopus subject areas

  • Biochemistry

Cite this

ATP-binding Cassette Transporter A1 Contains an NH2-terminal Signal Anchor Sequence That Translocates the Protein's First Hydrophilic Domain to the Exoplasmic Space. / Fitzgerald, Michael L.; Mendez, Armando J; Moore, Kathryn J.; Andersson, Lorna P.; Panjeton, Hess A.; Freeman, Mason W.

In: Journal of Biological Chemistry, Vol. 276, No. 18, 04.05.2001, p. 15137-15145.

Research output: Contribution to journalArticle

Fitzgerald, Michael L. ; Mendez, Armando J ; Moore, Kathryn J. ; Andersson, Lorna P. ; Panjeton, Hess A. ; Freeman, Mason W. / ATP-binding Cassette Transporter A1 Contains an NH2-terminal Signal Anchor Sequence That Translocates the Protein's First Hydrophilic Domain to the Exoplasmic Space. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 18. pp. 15137-15145.
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