title: L-Plastin S-glutathionylation promotes reduced binding to β-actin and affects neutrophil functions.
abstract: Posttranslational modifications (PTMs) of cytoskeleton proteins due to oxidative stress associated with several pathological conditions often lead to alterations in cell function. The current study evaluates the effect of nitric oxide (DETA-NO)-induced oxidative stress-related S-glutathionylation of cytoskeleton proteins in human PMNs. By using in vitro and genetic approaches, we showed that S-glutathionylation of L-plastin (LPL) and β-actin promotes reduced chemotaxis, polarization, bactericidal activity, and phagocytosis. We identified Cys-206, Cys-283, and Cys-460as S-thiolated residues in the β-actin-binding domain of LPL, where cys-460 had the maximum score. Site-directed mutagenesis of LPL Cys-460 further confirmed the role in the redox regulation of LPL. S-Thiolation diminished binding as well as the bundling activity of LPL. The presence of S-thiolated LPL was detected in neutrophils from both diabetic patients and db/db mice with impaired PMN functions. Thus, enhanced nitroxidative stress may results in LPL S-glutathionylation leading to impaired chemotaxis, polarization, and bactericidal activity of human PMNs, providing a mechanistic basis for their impaired functions in diabetes mellitus.
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Megha
authors.initials: M
authors.lastname: Dubey
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Abhishek K
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Deepika
authors.initials: D
authors.lastname: Awasthi
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Sheela
authors.initials: S
authors.lastname: Nagarkoti
authors.affiliation: Division of Experimental Hematology and Cancer Biology, Cincinnati Children׳s Research Foundation, Cincinnati, OH 45229, USA.
authors.forename: Sachin
authors.initials: S
authors.lastname: Kumar
authors.affiliation: King George׳s Medical University, Lucknow, India.
authors.forename: Wahid
authors.initials: W
authors.lastname: Ali
authors.affiliation: King George׳s Medical University, Lucknow, India.
authors.forename: Tulika
authors.initials: T
authors.lastname: Chandra
authors.affiliation: Centre for Cellular and Molecular Platforms, National Centre for Biological Sciences (NCBS-TIFR), Bangalore, India.
authors.forename: Vikas
authors.initials: V
authors.lastname: Kumar
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Manoj K
authors.initials: MK
authors.lastname: Barthwal
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Kumaravelu
authors.initials: K
authors.lastname: Jagavelu
authors.affiliation: Centro de Biología Molecular 'Severo Ochoa' (CSIC-UAM), Campus Universidad Autónoma, Nicolás, Cabrera 1, E-28049, Madrid, Spain.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Sánchez-Gómez
authors.affiliation: Centro de Biología Molecular 'Severo Ochoa' (CSIC-UAM), Campus Universidad Autónoma, Nicolás, Cabrera 1, E-28049, Madrid, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Lamas
authors.affiliation: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India. Electronic address: madhu_dikshit@cdri.res.in.
authors.forename: Madhu
authors.initials: M
authors.lastname: Dikshit
mesh_terms: D000199:Actins / Q000378:metabolism*; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D016022:Case-Control Studies; D016764:Cell Polarity; D002633:Chemotaxis; D003920:Diabetes Mellitus / Q000276:immunology / Q000378:metabolism; D005260:Female; D005978:Glutathione / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008810:Mice, Inbred C57BL; D008820:Mice, Obese; D008840:Microfilament Proteins / Q000378:metabolism*; D008875:Middle Aged; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism; D018384:Oxidative Stress; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D055815:Young Adult
chemical_list: D000199:Actins; C047262:LCP1 protein, human; D008840:Microfilament Proteins; D009569:Nitric Oxide; D005978:Glutathione
keywords: Bactericidal activity; Chemotaxis; Diabetes; Nitric oxide; Oxidative stress; Phagocytosis; Polarization; Posttranslational modifications