Interaction of the N-terminus of chicken skeletal essential light chain 1 with F-actin

O. A. Andreev, L. D. Saraswat, S. Lowey, C. Slaughter, J. Borejdo

Research output: Contribution to journalArticle

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Abstract

Skeletal myosin has two isoforms of the essential light chain (ELC), called LC1 and LC3, which differ only in their N-terminal amino acid sequence. The LC1 has 41 additional residues containing seven pairs of Ala- Pro, which form an elongated structure, and two pairs of lysines located near the N-terminus. When myosin subfragment-1 (S1) binds to actin, these lysines may interact with the C-terminus of actin and be responsible for the isoform specific properties of myosin. Here we employ cross-linking to identify the LC1 residues that are in contact with actin. S1 was reconstituted with various LC1 mutants and reacted with the zero-length cross-linker 1-ethyl-3- [3-dimethyl-aminopropyl]-carbodiimide (EDC). Cross-linking occurred only when actin was in molar excess over S1. Wild-type LC1 could be cross-linked through the terminal α-NH2 group, as well as via the two pairs of lysines. In a mutant ELC, where the lysines were deleted but two arginines were introduced near the N-terminus, the light chain could still be cross-linked via the terminal α-NH2 group. When the charge was reduced in the N-terminal region while retaining the Ala-Pro rich region, the mutant could not be cross-linked. These results suggest that as long as the N-terminus contains charged residues and an Ala-Pro rich extension, the binding between LC1 and actin can occur.

Original languageEnglish (US)
Pages (from-to)2480-2485
Number of pages6
JournalBiochemistry
Volume38
Issue number8
DOIs
StatePublished - Feb 23 1999

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alanylproline
Actins
Chickens
Lysine
Light
Myosins
Protein Isoforms
Myosin Subfragments
Arginine
Amino Acid Sequence
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interaction of the N-terminus of chicken skeletal essential light chain 1 with F-actin. / Andreev, O. A.; Saraswat, L. D.; Lowey, S.; Slaughter, C.; Borejdo, J.

In: Biochemistry, Vol. 38, No. 8, 23.02.1999, p. 2480-2485.

Research output: Contribution to journalArticle

Andreev, O. A. ; Saraswat, L. D. ; Lowey, S. ; Slaughter, C. ; Borejdo, J. / Interaction of the N-terminus of chicken skeletal essential light chain 1 with F-actin. In: Biochemistry. 1999 ; Vol. 38, No. 8. pp. 2480-2485.
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