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Updated: Oct 24, 2026

Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP)
Published on: April 20, 2015
A membrane protein primarily associated with the lysosomal compartment
D A Brooks1, T M Bradford, S R Carlsson
1Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide, SA, Australia. dbrooks@medicine.adelaide.edu.au
Abstract:
A monoclonal antibody designated MBR 39 has been generated against a membrane associated protein found selectively on lysosomes. MBR 39 reacts with the cytosolic face of the lysosome and was used to develop an organelle binding assay which reacted with high density organelles characteristic of lysosomes. These organelles contained lysosomal enzyme markers which included the integral membrane protein acetyl-CoA:alpha-glucosaminide N-acetyltransferase and the soluble lysosomal enzyme markers acid phosphatase (mature form), beta-hexosaminidase, arylsulfatase, and alpha-L-iduronidase. Under conditions which disrupt lysosomes the release of the latter soluble lysosomal enzymes was demonstrated from MBR 39 bound organelles. Immunoblots of MBR 39 with purified fibroblast lysosomal membrane, demonstrated reactivity with polypeptides of molecular mass 63 kDa (major species) and 73 kDa (minor species).
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