title: Structure-Function Analysis of the Non-Muscle Myosin Light Chain Kinase (nmMLCK) Isoform by NMR Spectroscopy and Molecular Modeling: Influence of MYLK Variants.
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.
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Kui
authors.initials: K
authors.lastname: Shen
authors.affiliation: Center for Structural Biology, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Ramirez
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Brandon
authors.initials: B
authors.lastname: Mapes
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Grace R
authors.initials: GR
authors.lastname: Shen
authors.affiliation: College of Pharmacy and BIO5 Institute, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Vijay
authors.initials: V
authors.lastname: Gokhale
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Mary E
authors.initials: ME
authors.lastname: Brown
authors.affiliation: Center for Structural Biology, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Bernard
authors.initials: B
authors.lastname: Santarsiero
authors.affiliation: Center for Structural Biology, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ishii
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Dudek
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America; Arizona Respiratory Center and Department of Medicine, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Institute for Personalized Respiratory Medicine, Section of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, Chicago, Illinois, United States of America; Arizona Respiratory Center and Department of Medicine, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Joe G N
authors.initials: JG
authors.lastname: Garcia
mesh_terms: D044383:Black People / Q000235:genetics; D006801:Humans; D008171:Lung Diseases / Q000235:genetics* / Q000473:pathology; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D020033:Protein Isoforms; D009219:Myosin-Light-Chain Kinase; C000593292:non-muscle myosin light-chain kinase, human