Coupled proteolytic and mass spectrometry studies indicate a novel topology for the glycine receptor

Citation for published article

Issues and recommendations for exploratory factor analysis and principal component analysis

DOI

10.1074/jbc.275.18.13683

Document Type

Journal Article

Publication Date

5-5-2000

Publication Title

Journal of Biological Chemistry

School

Bayer School of Natural and Environmental Sciences

Abstract

Members of the heteropentameric ligand-gated ion channel superfamily rapidly mediate signaling across the synaptic cleft. Sequence analysis and limited experimental studies have yielded a topological model containing four transmembrane α-helices, labeled M1 to M4, and a large soluble, extracellular N-terminal domain. This model persists to date despite some recent structural studies that suggest it may be inappropriate. In this study, the topology of the glycine receptor was probed by limited proteolysis coupled to mass spectrometry. Of particular note, accessible cleavage sites within the putative M1 and M3 transmembrane helices were identified. Membrane-associated fragments within the postulated globular extracellular N- terminal domain were also observed. This report presents several key details incorporated in a new topological model and is the first direct experimental evidence that a subset of the transmembrane regions are too short to be membrane-spanning α-helices; rather, these regions are proposed to be a mix of α-helices and β-sheets. This report is also the first to exploit the capability of mass spectrometry to probe critically the topology of a class of membrane proteins of unknown structure.

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