Measurement of glycosylated amino acids and peptides from urine of diabetic patients using affinity chromatography

M. Brownlee, H. Vlassara, A. Cerami

Research output: Contribution to journalArticle

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Abstract

An affinity chromatography system has been developed that retains glycosylated amino acids and peptides. Using this system, synthetic 14C-glycosylated lysine (reduced with NaB3H4) was completely separated from a mixture of reduced 14C-glycosylated lysine and unmodified 3H-lysine. Amino acid analysis of the retained peak from hydrolyzed human diabetic hemoglobin previously reduced with NaB3H4 revealed an equimolar mixture of glycosylated valine and glycosylated lysine, in agreement with previously published data obtained using other methodologies. These data demonstrate that in alkaline solution, the NaB3H4-reducible breakdown products of nonenzymatically glycosylated proteins are adsorbed to m-aminophenyl boronic acid immobilized on Bio-Gel P-6, while nonglycosylated amino acids are not. This affinity chromatography system should facilitate the rapid evaluation of nonenzymatic glycosylation in a large number of diabetic tissues. Levels of retained compounds in urine from diabetic and normal patients were determined by measuring ninhydrin-positive material. Amino acid analysis of NaB3H4-reduced hydrolysates of these peaks showed that glycosylated lysine was the major borohydride-reducible adduct present (67%-86%). Linear regression analysis showed that the quantity of excreted compounds in normals correlated with body weight (r = 0.63). The mean level (μmol leucine-equivalent/24 hr/kg body weight) in diabetics was over 1.5 times that found in urine from normal subjects (P < 0.005).

Original languageEnglish (US)
Pages (from-to)1044-1047
Number of pages4
JournalDiabetes
Volume29
Issue number12
StatePublished - 1980
Externally publishedYes

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Affinity Chromatography
Urine
Amino Acids
Peptides
Body Weight
Boronic Acids
Ninhydrin
Borohydrides
Valine
Glycosylation
Leucine
Lysine
Linear Models
Hemoglobins
Gels
Regression Analysis
glycosylated lysine
Proteins

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Internal Medicine

Cite this

Measurement of glycosylated amino acids and peptides from urine of diabetic patients using affinity chromatography. / Brownlee, M.; Vlassara, H.; Cerami, A.

In: Diabetes, Vol. 29, No. 12, 1980, p. 1044-1047.

Research output: Contribution to journalArticle

Brownlee, M. ; Vlassara, H. ; Cerami, A. / Measurement of glycosylated amino acids and peptides from urine of diabetic patients using affinity chromatography. In: Diabetes. 1980 ; Vol. 29, No. 12. pp. 1044-1047.
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