title: Human acetyl-CoA:glucosamine-6-phosphate N-acetyltransferase 1 has a relaxed donor specificity and transfers acyl groups up to four carbons in length.
abstract: Glucosamine-6-phosphate N-acetyltransferase1 (GNA1) catalyses the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to glucosamine-6-phosphate (GlcN6P) to form N-acetylglucosamine-6-phosphate (GlcNAc6P), which is an essential intermediate in UDP-GlcNAc biosynthesis. An analog of GlcNAc, N-butyrylglucosamine (GlcNBu) has shown healing properties for bone and articular cartilage in animal models of arthritis. The goal of this work was to examine whether GNA1 has the ability to transfer a butyryl group from butyryl-CoA to GlcN6P to form GlcNBu6P, which can then be converted to GlcNBu. We developed fluorescent and radioactive assays and examined the donor specificity of human GNA1. Acetyl, propionyl, n-butyryl, and isobutyryl groups were all transferred to GlcN6P, but isovaleryl-CoA and decanoyl-CoA did not serve as donor substrates. Site-specific mutants were produced to examine the role of amino acids potentially affecting the size and properties of the AcCoA binding pocket. All of the wild type and mutant enzymes showed activities of both acetyl and butyryl transfer and can therefore be used for the enzymatic synthesis of GlcNBu for biomedical applications.
authors.affiliation: a Department of Medicine, Division of Rheumatology, Queen's University, Kingston, ON K7L 3N6, Canada.|b Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada.
authors.forename: Inka
authors.initials: I
authors.lastname: Brockhausen
authors.affiliation: a Department of Medicine, Division of Rheumatology, Queen's University, Kingston, ON K7L 3N6, Canada.|c Ministry of Higher Education, Sur College of Applied Sciences, Sur, Sultanate of Oman.
authors.forename: Dileep G
authors.initials: DG
authors.lastname: Nair
authors.affiliation: b Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada.|d School of Life Sciences and The State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: b Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Yang
authors.affiliation: b Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada.
authors.forename: John S
authors.initials: JS
authors.lastname: Allingham
authors.affiliation: e Department of Chemistry, Queen's University, Kingston, ON K7L 3N6, Canada.
authors.forename: Walter A
authors.initials: WA
authors.lastname: Szarek
authors.affiliation: a Department of Medicine, Division of Rheumatology, Queen's University, Kingston, ON K7L 3N6, Canada.|b Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON K7L 3N6, Canada.
authors.forename: Tassos
authors.initials: T
authors.lastname: Anastassiades
mesh_terms: D000105:Acetyl Coenzyme A / Q000737:chemistry / Q000378:metabolism*; D002244:Carbon / Q000737:chemistry / Q000378:metabolism*; D005453:Fluorescence; D005944:Glucosamine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000737:chemistry; D051081:Glucosamine 6-Phosphate N-Acetyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019298:Glucose-6-Phosphate / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000737:chemistry; D006801:Humans; D013053:Spectrophotometry
chemical_list: C001293:glucosamine 6-phosphate; D019298:Glucose-6-Phosphate; D000105:Acetyl Coenzyme A; D002244:Carbon; C494970:GNPNAT1 protein, human; D051081:Glucosamine 6-Phosphate N-Acetyltransferase; D005944:Glucosamine
keywords: GlcNBu; HsGNA1; acetyl-CoA binding; butyryl-CoA; dosage en radioactivité; dosage par fluorescence; fluorescence assays; liaison de l’acétyl-CoA; mutagenèse dirigée; radioactive assays; site-directed mutagenesis