Characterization of bovine serum albumin glycated with glucose, galactose and lactose.
The non-enzymatic reaction between reducing sugars and proteins, known as glycation, has received increased attention from nutritional and medical research. In addition, there is a large interest in obtaining glycoconjugates of pure well-characterized oligosaccharides for biological research. In this study, glycation of bovine serum albumin (BSA) by d-glucose, d-galactose and d-lactose under dry-heat at 60 degrees C for 30, 60, 120, 180 or 240 min was assessed and the glycated products studied in order to establish their biological recognition by lectins. BSA glycation was monitored using gel electrophoresis, determination of available amino groups and lectin binding assays. The BSA molecular mass increase and glycation sites were investigated by mass spectrometry and through digestion with trypsin and chymotrypsin. Depending on time and type of sugar, differences in BSA conjugation were achieved. Modified BSA revealed reduction of amino groups' availability and slower migration through SDS/PAGE. d-galactose was more reactive than d-glucose or d-lactose, leading to the coupling of 10, 3 and 1 sugar residues, respectively, after 120 minutes of reaction. BSA lysines (K) were the preferred modified amino acids; both K256 and K420 appeared the most available for conjugation. Only BSA-lactose showed biological recognition by specific lectins.
Ledesma-Osuna
Ana Irene
AI
Centro de Investigación en Alimentación y Desarrollo, Sonora, México.
Ramos-Clamont
Gabriela
G
Vázquez-Moreno
Luz
L
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005690:Galactose / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D017127:Glycation End Products, Advanced; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D006358:Hot Temperature; D066298:In Vitro Techniques; D007785:Lactose / Q000737:chemistry; D037102:Lectins / Q000378:metabolism; D015416:Maillard Reaction; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000093423:Glycated Serum Albumin
D017127:Glycation End Products, Advanced; D006023:Glycoproteins; D037102:Lectins; D010446:Peptide Fragments; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D005947:Glucose; D007785:Lactose; D005690:Galactose; D000093423:Glycated Serum Albumin