Outer membrane monolayer domains from two-dimensional surface scanning resistance measurements

Kenichi Suzuki, Ronald E. Sterba, Michael Sheetz

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

Cellular plasma membranes have domains that are defined, in most cases, by cytoskeletal elements. The outer half of the bilayer may also contain domains that organize glycosylphosphatidylinositol (GPI)-linked proteins. To define outer membrane barriers, we measured the resistive force on membrane bound beads as they were scanned across the plasma membrane of HEPA-OVA cells with optical laser tweezers. Beads were bound by antibodies to fluorescein- phosphatidylethanolamine (FI-PE) or to the class I major histocompatibility complex (MHC class I) Qa-2 (a GPI-anchored protein). Two-dimensional scans of resistive force showed both occasional, resistive barriers and a velocity- dependent, continuous resistance. At the lowest antibody concentration, which gave specific binding, the continuous friction coefficient of Qa-2 was consistent with that observed by single-particle tracking (SPT) of small gold particles. At high antibody concentrations, the friction coefficient was significantly higher but decreased with increasing temperature, addition of deoxycholic acid, or treatment with heparinase I. Barriers to lateral movement (>3 times the continuous resistance) were consistently observed. Elastic barriers (with elastic constants from 1 to 20 pN/μm and sensitive to cytochalasin D) and small nonelastic barriers (< 100 nm) were specifically observed with beads bound to the GPI-linked Qa-2. We suggest that GPI-linked proteins interact with transmembrane proteins when aggregated by antibody- coated beads and the transmembrane proteins encounter cytoplasmic barriers to lateral movement. The barriers to lateral movement are dynamic, discontinuous, and low in density.

Original languageEnglish (US)
Pages (from-to)448-459
Number of pages12
JournalBiophysical Journal
Volume79
Issue number1
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics

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