Chemical cross-linking indicates a staggered and antiparallel protofilament of desmin intermediate filaments and characterizes one higher-level complex between protofilaments

Authors

  • Norbert GEISLER,

    Corresponding author
    1. Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Göttingen, Federal Republic of Germany
      Correspondence to N. Geisler, Max Planck Institute for Biophysical Chemistry, W-3400 Göttingen, P.O. Box 2841, Federal Republic of Germany
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  • Jürgen SCHÜNEMANN,

    1. Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Göttingen, Federal Republic of Germany
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  • Klaus WEBER

    1. Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Göttingen, Federal Republic of Germany
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Correspondence to N. Geisler, Max Planck Institute for Biophysical Chemistry, W-3400 Göttingen, P.O. Box 2841, Federal Republic of Germany

Abstract

Tetrameric rods, protofilaments and assembled filaments of desmin, the intermediate filament protein of muscle, have been chemically cross-linked with the lysine specific cross-linkers EGS [ethylene glycol bis(succinimidylsuccinate), 1.61 nm span] and bis(sulfosuccinimidyl) suberate (1.14 nm span). One bis(sulfosuccinimidyl)suberate and two EGS cross-links were isolated from the rod and characterized. They show that the two coiled coils in the rod tetramer are staggered by approximately 15–20 nm and strongly indicate an antiparallel arrangement in which the inner overlapping part of the rod is formed by the amino-terminal helices 1A, 1B and 2A. Both EGS cross-links identified in the rod were also isolated from cross-linked filaments. The isolated rod, therefore, represents a complex also present in identical, or very similar form in protofilaments and in assembled filaments. Cross-linked filaments yielded a third EGS cross-link that must have been formed between neighboring protofilaments. It connects the highly conserved carboxy-terminus of helix 2B of the first protofilament to the overlap region formed by helices 1A and 2A of the second protofilament. The restrictions posed by these cross-links on current filament models are discussed.

Abbreviations
EGS

ethylene glycol bis(succinimidylsuccinate)

IF

intermediate filament