title: Proteomic profiling and post-translational modifications in human keratinocytes treated with Mucuna pruriens leaf extract.
abstract: BACKGROUND
  Mucuna pruriens (Mp) is a plant belonging to the Fabaceae family, with several medicinal properties among which its potential to treat diseases where reactive oxygen species (ROS) play an important role in the pathogeneses. The aim was to investigate the effects of Mp leaf methanolic extract (MPME) on human keratinocytes protein expression and its role in preventing proteins oxidation after oxidative stress (OS) exposure.
  
  
  METHODS
  The effects of MPME on HaCaT cells protein expression were evaluated treating cells with different concentrations of MPME, with glucose oxidase (GO, source of OS) and with MPME subsequently treated with GO. The protein patterns of treated HaCaT cells are analyzed by two-dimensional gel electrophoresis (2-DE) and compared with that of untreated HaCaT. Immunoblotting was then used to evaluate the role of MPME in preventing the 4-hydroxynonenal protein adducts (4-HNE PAs) formation (marker of OS).
  
  
  RESULTS
  Eighteen proteins, identified by mass spectrometry (LC-ESI-CID-MS/MS), were modulated distinctly by MPME in HaCaT. Overall, MPME counteract GO effect, reducing the GO-induced overexpression of several proteins involved in stress response (T-complex protein 1, Protein disulfide-isomerase A3, Protein DJ-1, and Stress-induced-phosphoprotein 1), in cell energy methabolism (Inorganic pyrophosphatase, Triosephosphate isomerase isoform 1, 2-phosphopyruvate-hydratase alpha-enolase, and Fructose-bisphosphate aldolase A isoform 1), in cytoskeletal organization (Cytokeratins 18, 9, 2, Cofilin-1, Annexin A2 and F-actin-capping protein subunit beta isoform 1) and in cell cycle progression (Eukaryotic translation initiation factor 5A-1 isoform B). In addition, MPME decreased the 4-HNE PAs levels, in particular on 2-phosphopyruvate-hydratase alpha-enolase and Cytokeratin 9.
  
  
  CONCLUSIONS
  Our findings show that MPME might be helpful in the treatment of OS-related skin diseases by preventing protein post-translational modifications (4-HNE PAs).
authors: Department of Medical Biotechnologies, University of Siena, Siena, Italy; Child Neuropsychiatry Unit, University Hospital AOUS, Siena, Italy.
authors: Alessio
authors: A
authors: Cortelazzo
authors: Department of Biotechnology, Chemistry and Pharmaceutical Sciences, University of Siena, Siena, Italy.
authors: Raffaella L
authors: RL
authors: Lampariello
authors: Department of Life Sciences and Biotechnologies, University of Ferrara, Via Luigi Borsari 46, 44100 Ferrara, Italy.
authors: Claudia
authors: C
authors: Sticozzi
authors: Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors: Roberto
authors: R
authors: Guerranti
authors: Department of Ecological and Biological Sciences, University of Tuscia, Viterbo, Italy.
authors: Cristiana
authors: C
authors: Mirasole
authors: Department of Ecological and Biological Sciences, University of Tuscia, Viterbo, Italy.
authors: Lello
authors: L
authors: Zolla
authors: Department of Life Sciences and Biotechnologies, University of Ferrara, Via Luigi Borsari 46, 44100 Ferrara, Italy.
authors: Gianni
authors: G
authors: Sacchetti
authors: Child Neuropsychiatry Unit, University Hospital AOUS, Siena, Italy.
authors: Joussef
authors: J
authors: Hajek
authors: Department of Life Sciences and Biotechnologies, University of Ferrara, Via Luigi Borsari 46, 44100 Ferrara, Italy; Department of Food and Nutrition, Kyung Hee University, Seoul, Rrepbulic of Korea. Electronic address: giuseppe.valacchi@unife.it.
authors: Giuseppe
authors: G
authors: Valacchi
mesh_terms: D000447:Aldehydes / Q000378:metabolism; D002460:Cell Line; D005949:Glucose Oxidase / Q000494:pharmacology; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D031299:Mucuna; D018384:Oxidative Stress; D010936:Plant Extracts / Q000494:pharmacology*; D018515:Plant Leaves; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011506:Proteins / Q000378:metabolism; D040901:Proteomics
chemical_list: D000447:Aldehydes; D010936:Plant Extracts; D011506:Proteins; D005949:Glucose Oxidase; C027576:4-hydroxy-2-nonenal
keywords: Keratinocytes; Leaf extract; Mucuna pruriens; Oxidative stress; Proteomics