N-glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C-CAM, from rat liver: detection of oversialylated bi- and triantennary structures.
Rat C-CAM is a ubiquitous, transmembrane and carcinoembryonic antigen related cell adhesion molecule. The human counterpart is known as biliary glycoprotein (BGP) or CD66a. It is involved in different cellular functions ranging from intercellular adhesion, microbial receptor activity, signaling and tumor suppression. In the present study N-glycosylation of C-CAM immunopurified from rat liver was analyzed in detail. The primary sequence of rat C-CAM contains 15 potential N-glycosylation sites. The N-glycans were enzymatically released from glycopeptides, fluorescently labeled with 2-aminobenzamide, and separated by two-dimensional HPLC. Oligosaccharide structures were characterized by enzymatic sequencing and MALDI-TOF-MS. Mainly bi- and triantennary complex structures were identified. The presence of type I and type II chains in the antennae of these glycans results in heterogeneous glycosylation of C-CAM. Sialylation of the sugars was found to be unusual; bi- and triantennary glycans contained three and four sialic acid residues, respectively, and this linkage seemed to be restricted to the type I chain in the antennae. Approximately 20% of the detected sugars contain these unusual numbers of sialic acids. C-CAM is the first transmembrane protein found to be oversialylated.
Kannicht
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Institut für Molekularbiologie und Biochemie der Freien Universität Berlin, Fachbereich Humanmedizin, Berlin-Dahlem, Germany.
Lucka
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L
Nuck
R
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Reutter
W
W
Gohlke
M
M
D000251:Adenosine Triphosphatases / Q000737:chemistry*; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry; D000943:Antigens, Differentiation / Q000737:chemistry; D002240:Carbohydrate Sequence; D002272:Carcinoembryonic Antigen / Q000737:chemistry*; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D008099:Liver / Q000737:chemistry*; D008297:Male; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D017421:Sequence Analysis; D012795:Sialoglycoproteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015703:Antigens, CD; D000943:Antigens, Differentiation; C069847:CD66 antigens; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D009844:Oligosaccharides; D012795:Sialoglycoproteins; D000251:Adenosine Triphosphatases