An assay to detect glycoproteins that contain mannose 6-phosphate

Kenneth J. Valenzano, Laura M. Kallay, Peter Lobel

Research output: Contribution to journalArticlepeer-review

41 Scopus citations

Abstract

We have developed a broadly applicable solid phase assay for quantitative and qualitative analysis of phosphorylated lysosomal enzymes and other glycoproteins that contain mannose 6-phosphate (Man6-P) residues. Samples are immobilized on membranes and proteins that contain Man6-P are detected using an iodinated, soluble form of the cation-independent Man6-P receptor. The assay is highly specific, sensitive, and linear over 3 orders of magnitude. The technique has two general applications. First, it can be used in a dot blot format to analyze large numbers of samples for total levels of Man6-P glycoproteins. Second, it can be used in a western blot format to identify Man6-P glycoproteins that have been fractionated by polyacrylamide gel electrophoresis. Specific applications of this novel assay include structural characterization of glycoproteins, diagnosis of mucolipidosis II and III, and screening of pharmaceuticals for lysosomotropic activity. Other potential applications are discussed.

Original languageEnglish (US)
Pages (from-to)156-162
Number of pages7
JournalAnalytical Biochemistry
Volume209
Issue number1
DOIs
StatePublished - Feb 15 1993

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biophysics
  • Biochemistry
  • Cell Biology

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