title: Click-chemistry approach to study mycoloylated proteins: Evidence for PorB and PorC porins mycoloylation in Corynebacterium glutamicum.
abstract: Protein mycoloylation is a recently identified, new form of protein acylation. This post-translational modification consists in the covalent attachment of mycolic acids residues to serine. Mycolic acids are long chain, α-branched, β-hydroxylated fatty acids that are exclusively found in the cell envelope of Corynebacteriales, a bacterial order that includes important genera such as Mycobacterium, Nocardia or Corynebacterium. So far, only 3 mycoloylated proteins have been identified: PorA, PorH and ProtX from C. glutamicum. Whereas the identity and function of ProtX is unknown, PorH and PorA associate to form a membrane channel, the activity of which is dependent upon PorA mycoloylation. However, the exact role of mycoloylation and the generality of this phenomenon are still unknown. In particular, the identity of other mycoloylated proteins, if any, needs to be determined together with establishing whether such modification occurs in Corynebacteriales genera other than Corynebacterium. Here, we tested whether a metabolic labeling and click-chemistry approach could be used to detect mycoloylated proteins. Using a fatty acid alkyne analogue, we could indeed label PorA, PorH and ProtX and determine ProtX mycoloylation site. Importantly, we also show that two other porins from C. glutamicum, PorB and PorC are mycoloylated.
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.|Holy Spirit University of Kaslik (USEK), Jounieh, Mount Lebanon, Lebanon.
authors.forename: Hanane
authors.initials: H
authors.lastname: Issa
authors.affiliation: Institute of Pharmacology and Structural Biology (IPBS), UMR 5089, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Huc-Claustre
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.
authors.forename: Thamila
authors.initials: T
authors.lastname: Reddad
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.
authors.forename: Nolwenn
authors.initials: N
authors.lastname: Bonadé Bottino
authors.affiliation: Institute of Pharmacology and Structural Biology (IPBS), UMR 5089, France.
authors.forename: Maryelle
authors.initials: M
authors.lastname: Tropis
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Houssin
authors.affiliation: Institute of Pharmacology and Structural Biology (IPBS), UMR 5089, France.
authors.forename: Mamadou
authors.initials: M
authors.lastname: Daffé
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bayan
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette Cedex, France.
authors.forename: Nathalie
authors.identifier: http://orcid.org/0000-0003-1765-4003
authors.initials: N
authors.lastname: Dautin
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D057930:Click Chemistry; D048230:Corynebacterium glutamicum / Q000378:metabolism*; D005227:Fatty Acids / Q000737:chemistry; D009171:Mycolic Acids / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D018272:Porins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D009171:Mycolic Acids; D018272:Porins