Research ExplorerPublicationsThe binding of DYNLL2 to myosin Va requires alternatively sp...

The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain

Standard

The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain. / Wagner, Wolfgang; Fodor, Elfrieda; Ginsburg, Ann; Hammer, John A.

In: BIOCHEMISTRY-US, Vol. 45, No. 38, 26.09.2006, p. 11564-77.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Wagner, W, Fodor, E, Ginsburg, A & Hammer, JA 2006, 'The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain', BIOCHEMISTRY-US, vol. 45, no. 38, pp. 11564-77. https://doi.org/10.1021/bi061142u

APA

Wagner, W., Fodor, E., Ginsburg, A., & Hammer, J. A. (2006). The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain. BIOCHEMISTRY-US, 45(38), 11564-77. https://doi.org/10.1021/bi061142u

Vancouver

Wagner W, Fodor E, Ginsburg A, Hammer JA. The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain. BIOCHEMISTRY-US. 2006 Sep 26;45(38):11564-77. https://doi.org/10.1021/bi061142u

Bibtex

@article{f82feb76ada84b6c86ef2d9f854e1db7,
title = "The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain",
abstract = "The myosin Va light chain DYNLL2 has been proposed to function as an adaptor to link the myosin to certain cargo. Here, we mapped the binding site for DYNLL2 within the myosin Va heavy chain. Copurification and pull-down experiments showed that the heavy chain contains a single DYNLL2 binding site and that this site resides within a discontinuity in the myosin's central coiled-coil domain. Importantly, exon B, an alternatively spliced, three-amino acid exon, is a part of this binding site, and we show in the context of full-length myosin Va that this exon is required for DYNLL2-myosin Va interaction. We investigated the effect of DYNLL2 binding on the structure of a myosin Va heavy chain fragment that contains the DYNLL2 binding site and flanking sequence, only parts of which are strongly predicted to form a coiled coil. Circular dichroism measurements revealed a DYNLL2-induced change in the secondary structure of this dimeric myosin fragment that is consistent with an increase in alpha-helical coiled-coil content. Moreover, the binding of DYNLL2 considerably stabilizes this heavy chain fragment against thermal denaturation. Analytical ultracentrifugation yielded an apparent association constant of approximately 3 x 10(6) M(-1) for the interaction of DYNLL2 with the dimeric myosin fragment. Together, these data show that alternative splicing of the myosin Va heavy chain controls DYNLL2-myosin Va interaction and that DYNLL2 binding alters the structure of a portion of the myosin's coiled-coil domain. These results suggest that exon B could have a significant impact on the conformation and regulatory folding of native myosin Va, as well as on its interaction with certain cargos.",
keywords = "Alternative Splicing, Amino Acid Sequence, Amino Acids, Animals, Binding Sites, Circular Dichroism, Cytoplasmic Dyneins, Dimerization, Exons, Humans, Mice, Models, Biological, Molecular Sequence Data, Myosin Heavy Chains, Myosin Light Chains, Myosin Type V, Peptide Fragments, Protein Binding, Protein Folding, Protein Interaction Mapping, Protein Structure, Tertiary, Recombinant Fusion Proteins, Temperature, Thermodynamics",
author = "Wolfgang Wagner and Elfrieda Fodor and Ann Ginsburg and Hammer, {John A}",
year = "2006",
month = sep,
day = "26",
doi = "10.1021/bi061142u",
language = "English",
volume = "45",
pages = "11564--77",
journal = "BIOCHEMISTRY-US",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "38",

}

RIS

TY - JOUR

T1 - The binding of DYNLL2 to myosin Va requires alternatively spliced exon B and stabilizes a portion of the myosin's coiled-coil domain

AU - Wagner, Wolfgang

AU - Fodor, Elfrieda

AU - Ginsburg, Ann

AU - Hammer, John A

PY - 2006/9/26

Y1 - 2006/9/26

N2 - The myosin Va light chain DYNLL2 has been proposed to function as an adaptor to link the myosin to certain cargo. Here, we mapped the binding site for DYNLL2 within the myosin Va heavy chain. Copurification and pull-down experiments showed that the heavy chain contains a single DYNLL2 binding site and that this site resides within a discontinuity in the myosin's central coiled-coil domain. Importantly, exon B, an alternatively spliced, three-amino acid exon, is a part of this binding site, and we show in the context of full-length myosin Va that this exon is required for DYNLL2-myosin Va interaction. We investigated the effect of DYNLL2 binding on the structure of a myosin Va heavy chain fragment that contains the DYNLL2 binding site and flanking sequence, only parts of which are strongly predicted to form a coiled coil. Circular dichroism measurements revealed a DYNLL2-induced change in the secondary structure of this dimeric myosin fragment that is consistent with an increase in alpha-helical coiled-coil content. Moreover, the binding of DYNLL2 considerably stabilizes this heavy chain fragment against thermal denaturation. Analytical ultracentrifugation yielded an apparent association constant of approximately 3 x 10(6) M(-1) for the interaction of DYNLL2 with the dimeric myosin fragment. Together, these data show that alternative splicing of the myosin Va heavy chain controls DYNLL2-myosin Va interaction and that DYNLL2 binding alters the structure of a portion of the myosin's coiled-coil domain. These results suggest that exon B could have a significant impact on the conformation and regulatory folding of native myosin Va, as well as on its interaction with certain cargos.

AB - The myosin Va light chain DYNLL2 has been proposed to function as an adaptor to link the myosin to certain cargo. Here, we mapped the binding site for DYNLL2 within the myosin Va heavy chain. Copurification and pull-down experiments showed that the heavy chain contains a single DYNLL2 binding site and that this site resides within a discontinuity in the myosin's central coiled-coil domain. Importantly, exon B, an alternatively spliced, three-amino acid exon, is a part of this binding site, and we show in the context of full-length myosin Va that this exon is required for DYNLL2-myosin Va interaction. We investigated the effect of DYNLL2 binding on the structure of a myosin Va heavy chain fragment that contains the DYNLL2 binding site and flanking sequence, only parts of which are strongly predicted to form a coiled coil. Circular dichroism measurements revealed a DYNLL2-induced change in the secondary structure of this dimeric myosin fragment that is consistent with an increase in alpha-helical coiled-coil content. Moreover, the binding of DYNLL2 considerably stabilizes this heavy chain fragment against thermal denaturation. Analytical ultracentrifugation yielded an apparent association constant of approximately 3 x 10(6) M(-1) for the interaction of DYNLL2 with the dimeric myosin fragment. Together, these data show that alternative splicing of the myosin Va heavy chain controls DYNLL2-myosin Va interaction and that DYNLL2 binding alters the structure of a portion of the myosin's coiled-coil domain. These results suggest that exon B could have a significant impact on the conformation and regulatory folding of native myosin Va, as well as on its interaction with certain cargos.

KW - Alternative Splicing

KW - Amino Acid Sequence

KW - Amino Acids

KW - Animals

KW - Binding Sites

KW - Circular Dichroism

KW - Cytoplasmic Dyneins

KW - Dimerization

KW - Exons

KW - Humans

KW - Mice

KW - Models, Biological

KW - Molecular Sequence Data

KW - Myosin Heavy Chains

KW - Myosin Light Chains

KW - Myosin Type V

KW - Peptide Fragments

KW - Protein Binding

KW - Protein Folding

KW - Protein Interaction Mapping

KW - Protein Structure, Tertiary

KW - Recombinant Fusion Proteins

KW - Temperature

KW - Thermodynamics

U2 - 10.1021/bi061142u

DO - 10.1021/bi061142u

M3 - SCORING: Journal article

C2 - 16981716

VL - 45

SP - 11564

EP - 11577

JO - BIOCHEMISTRY-US

JF - BIOCHEMISTRY-US

SN - 0006-2960

IS - 38

ER -