Microheterogeneity of serum glycoproteins in patients with chronic alcohol abuse compared with carbohydrate-deficient glycoprotein syndrome type I.

BACKGROUND Chronic alcohol abuse alters the normal N-glycosylation of transferrin, producing the carbohydrate-deficient transferrin isoforms. This alteration could be similar to that present in patients with carbohydrate-deficient glycoprotein syndrome type 1 (CDG1). We thus compared the alterations of N-glycans present in patients with alcoholism and patients with CDG1. METHODS The N-glycans of serum glycoproteins were compared in sera of patients with alcoholism, patients with CDG1, and controls by two-dimensional electrophoresis, neuraminidase, peptide:N-glycosidase F, and endoglycosidase F2 treatments. A specific antibody directed against the amino acid sequence surrounding the N-432 N-glycosylation site of transferrin was prepared (SZ-350 antibody). RESULTS In patients with alcoholism, the abnormal transferrin and alpha(1)-antitrypsin isoforms were devoid of a variable number of entire N-glycan moieties and were identical with those present in CDG1. In the serum of patients with alcoholism, this finding was less pronounced than in CDG1. In contrast to CDG1, there was no decrease in clusterin or serum amyloid P in patients with alcoholism. The SZ-350 antibody recognized only transferrin isoforms with one or no N-glycan moieties. CONCLUSION Antibodies directed against specific N-glycosylation sites of glycoproteins could be useful for developing more specific immunochemical tests for the diagnosis of chronic alcohol abuse.

[1]  S. Borg,et al.  Evidence of a reduced sialic acid content in serum transferrin in male alcoholics. , 1981, Alcoholism, clinical and experimental research.

[2]  A. Görg,et al.  Two‐dimensional electrophoresis with immobilized pH gradients of leaf proteins from barley (Hordeum vulgare): Method, reproducibility and genetic aspects , 1988, Electrophoresis.

[3]  K. Yamashita,et al.  Electrospray ionization-mass spectrometric analysis of serum transferrin isoforms in patients with carbohydrate-deficient glycoprotein syndrome. , 1993, Journal of biochemistry.

[4]  D F Hochstrasser,et al.  Methods for increasing the resolution of two-dimensional protein electrophoresis. , 1988, Analytical biochemistry.

[5]  S. Borg,et al.  A modified method for the assay of carbohydrate-deficient transferrin (CDT) in serum. , 1991, Alcohol and alcoholism (Oxford, Oxfordshire). Supplement.

[6]  D. Hochstrasser,et al.  Micropreparative two‐dimensional electrophoresis allowing the separation of samples containing milligram amounts of proteins , 1993, Electrophoresis.

[7]  S. Borg,et al.  Micro anion exchange chromatography of carbohydrate-deficient transferrin in serum in relation to alcohol consumption (Swedish Patent 8400587-5). , 1986, Alcoholism, clinical and experimental research.

[8]  J. Mæhlen,et al.  [The carbohydrate deficient glycoprotein syndrome]. , 1991, Tidsskrift for den Norske laegeforening : tidsskrift for praktisk medicin, ny raekke.

[9]  S. Borg,et al.  Carbohydrate composition of serum transferrin in alcoholic patients. , 1986, Alcoholism, clinical and experimental research.

[10]  C. Lieber,et al.  Serum carbohydrate‐deficient transferrin: Mechanism of increase after chronic alcohol intake , 1995, Hepatology.

[11]  C. Seidel,et al.  Identification of carbohydrate deficient transferrin forms by MALDI-TOF mass spectrometry and lectin ELISABiochim Biophys Acta 1998 Aug 24;1381(3):356. , 1998, Biochimica et biophysica acta.

[12]  T. Arndt,et al.  Validation by isoelectric focusing of the anion-exchange isotransferrin fractionation step involved in determination of carbohydrate-deficient transferrin by the CDTect assay. , 1998, Clinical chemistry.

[13]  R. Bender,et al.  A General Regression Procedure for Method Transformation. Application of Linear Regression Procedures for Method Comparison Studies in Clinical Chemistry, Part III , 1988, Journal of clinical chemistry and clinical biochemistry. Zeitschrift fur klinische Chemie und klinische Biochemie.

[14]  M. Lakshman,et al.  Effects of chronic ethanol on enzymes regulating sialylation and desialylation of transferrin in rats. , 1993, Alcoholism, clinical and experimental research.

[15]  G. Beckman,et al.  Transferrin phenotype and level of carbohydrate-deficient transferrin in healthy individuals. , 1988, Alcoholism, clinical and experimental research.

[16]  J. Tissot,et al.  Microheterogeneity of serum glycoproteins and their liver precursors in patients with carbohydrate-deficient glycoprotein syndrome type I: apparent deficiencies in clusterin and serum amyloid P. , 1997, The Journal of laboratory and clinical medicine.

[17]  H. Stibler.  Direct immunofixation after isoelectric focusing An improved method for identification of cerebrospinal fluid and serum proteins , 1979, Journal of the Neurological Sciences.

[18]  M. Budarf,et al.  PMM (PMM1), the human homologue of SEC53 or yeast phosphomannomutase, is localized on chromosome 22q13. , 1997, Genomics.

[19]  E. Pardon,et al.  Mutations in PMM2, a phosphomannomutase gene on chromosome 16p13 in carbohydrate-deficient glycoprotein type I syndrome (Jaeken syndrome) , 1997, Nature Genetics.

[20]  A. Lundblad,et al.  Carbohydrate composition of serum transferrin isoforms from patients with high alcohol consumption. , 1995, Biochemical and biophysical research communications.

[21]  E. Bertini,et al.  Phosphomannose isomerase deficiency: a carbohydrate-deficient glycoprotein syndrome with hepatic-intestinal presentation. , 1998, American journal of human genetics.

[22]  Richard A. Lerner,et al.  Immunogenic structure of the influenza virus hemagglutinin , 1982, Cell.

[23]  D. Hochstrasser,et al.  New alterations of serum glycoproteins in alcoholic and cirrhotic patients revealed by high resolution two-dimensional gel electrophoresis. , 1996, Biochemical and biophysical research communications.

[24]  A. Hasilik,et al.  Elongation of N-acetyllactosamine repeats in diantennary oligosaccharides. , 1997, European journal of biochemistry.

[25]  J. C. Jamieson,et al.  The role of the carbohydrate chains of Galβ-1,4-GlcNAcα2,6-sialyltransferase for enzyme eactivity , 1993 .

[26]  C. Allgulander,et al.  Abnormal microheterogeneity of transferrin in serum and cerebrospinal fluid in alcoholism. , 2009, Acta medica Scandinavica.

[27]  E. Schaftingen,et al.  Phosphomannomutase deficiency is a cause of carbohydrate‐deficient glycoprotein syndrome type I , 1995, FEBS letters.