Structure-function analysis of the non- muscle myosin light chain kinase (nmMLCK) isoform by NMR spectroscopy and molecular modeling: Influence of MYLK variants

Kui Shen, Benjamin Ramirez, Brandon Mapes, Grace R. Shen, Vijay Gokhale, Mary E. Brown, Bernard Santarsiero, Yoshitaka Ishii, Steven M. Dudek, Ting Wang, Joe G N Garcia

Research output: Contribution to journalArticle

Abstract

The MYLK gene encodes the multifunctional enzyme, myosin light chain kinase (MLCK), involved in isoform-specific non-muscle and smooth muscle contraction and regulation of vascular permeability during inflammation. Three MYLK SNPs (P21H, S147P, V261A) alter the N-terminal amino acid sequence of the non-muscle isoform of MLCK (nmMLCK) and are highly associated with susceptibility to acute lung injury (ALI) and asthma, especially in individuals of African descent. To understand the functional effects of SNP associations, we examined the N-terminal segments of nmMLCK by 1H-15N heteronuclear single quantum correlation (HSQC) spectroscopy, a 2-D NMR technique, and by in silico molecular modeling. Both NMR analysis and molecular modeling indicated SNP localization to loops that connect the immunoglobulin-like domains of nmMLCK, consistent with minimal structural changes evoked by these SNPs. Molecular modeling analysis identified protein-protein interaction motifs adversely affected by these MYLK SNPs including binding by the scaffold protein 14-3-3, results confirmed by immunoprecipitation and western blot studies. These structure-function studies suggest novel mechanisms for nmMLCK regulation, which may confirm MYLK as a candidate gene in inflammatory lung disease and advance knowledge of the genetic underpinning of lung-related health disparities.

Original languageEnglish (US)
Article numbere0130515
JournalPLoS One
Volume10
Issue number6
DOIs
StatePublished - Jun 25 2015
Externally publishedYes

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myosin light chain kinase
Myosin-Light-Chain Kinase
Molecular modeling
Nuclear magnetic resonance spectroscopy
Single Nucleotide Polymorphism
Muscle
nuclear magnetic resonance spectroscopy
Protein Isoforms
Magnetic Resonance Spectroscopy
Muscles
muscles
Genes
lungs
Multifunctional Enzymes
Nuclear magnetic resonance
scaffolding proteins
Protein Interaction Domains and Motifs
14-3-3 Proteins
Pulmonary diseases
Acute Lung Injury

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Structure-function analysis of the non- muscle myosin light chain kinase (nmMLCK) isoform by NMR spectroscopy and molecular modeling : Influence of MYLK variants. / Shen, Kui; Ramirez, Benjamin; Mapes, Brandon; Shen, Grace R.; Gokhale, Vijay; Brown, Mary E.; Santarsiero, Bernard; Ishii, Yoshitaka; Dudek, Steven M.; Wang, Ting; Garcia, Joe G N.

In: PLoS One, Vol. 10, No. 6, e0130515, 25.06.2015.

Research output: Contribution to journalArticle

Shen, K, Ramirez, B, Mapes, B, Shen, GR, Gokhale, V, Brown, ME, Santarsiero, B, Ishii, Y, Dudek, SM, Wang, T & Garcia, JGN 2015, 'Structure-function analysis of the non- muscle myosin light chain kinase (nmMLCK) isoform by NMR spectroscopy and molecular modeling: Influence of MYLK variants', PLoS One, vol. 10, no. 6, e0130515. https://doi.org/10.1371/journal.pone.0130515
Shen, Kui ; Ramirez, Benjamin ; Mapes, Brandon ; Shen, Grace R. ; Gokhale, Vijay ; Brown, Mary E. ; Santarsiero, Bernard ; Ishii, Yoshitaka ; Dudek, Steven M. ; Wang, Ting ; Garcia, Joe G N. / Structure-function analysis of the non- muscle myosin light chain kinase (nmMLCK) isoform by NMR spectroscopy and molecular modeling : Influence of MYLK variants. In: PLoS One. 2015 ; Vol. 10, No. 6.
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