Gel Chromatographic Characterization of the Hydrophobic Interaction of Glycosylphosphatidylinositol-Alkaline Phosphatase with Detergents
-
Abstract
The interaction of a glycosylphosphatidylinositol (GPI) protein with different detergents was studied for the first time with a purified protein. Four differently hydrophobic fractions of GPIalkaline phosphatase (GPIAP) from calf intestine were used as model proteins. The mode of interaction was determined by investigating (i) the selfaggregation behaviour of the GPIAP fractions, (ii) the interference of detergents with GPIAP binding to octylSepharose, and (iii) the elution of GPIAP bound to octylSepharose. It was shown that polyoxyethylenetype detergents surprisingly interact much stronger than noctylglucoside with GPIAP, which is in contrast to the known behaviour of GPIproteins in natural membranes. Gel filtration chromatography of Triton X-100 at concentrations above the critical micellar concentration yields three different micelle species with apparent molecular weights of about 166, 54, and 16 kDa. GPIAP fraction II, which is shown to bear only one anchor per dimer, does not bind to any of these micelles. We demonstrate that a complex is formed containing about 150 Triton X-100 molecules and about 4700 molecules of water per molecule of GPIAP dimer. The experimental findings are in accordance with a simple geometrical model based on the physical data of fatty acids and the arrangement, mean size, and shape of Triton X-100 molecules.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- The Twin-Arginine Translocation System: A Novel Means of Transporting Folded Proteins in Chloroplasts and Bacteria
- De Novo DNA Methylation at Nonrandom Founder Sites 5' from an Unmethylated Minimal Origin of DNA Replication in Latent Epstein-Barr Virus Genomes
- Trichostatin A Modulates Expression of p21waf1/cip1, Bcl-xL, ID1, ID2, ID3, CRAB2, GATA-2, hsp86 and TFIID/TAFII31 mRNA in Human Lung Adenocarcinoma Cells
- Effects of the Active Aldehyde Group Generated by RNA N-Glycosidase in the Sarcin/Ricin Domain of Rat 28S Ribosomal RNA on Peptide Elongation
- Responses to Peroxynitrite in Yeast: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) as a Sensitive Intracellular Target for Nitration and Enhancement of Chaperone Expression and Ubiquitination
- Enhancing the T R Transition of Insulin by Helix-Promoting Sequence Modifications at the N-Terminal B-Chain
- Structure-Dependent Effects of Glucose-Containing Analogs of Platelet Activating Factor (PAF) on Membrane Integrity
- Two Different Mechanisms for Activation of Cyclic PIP Synthase: by a G Protein or by Protein Tyrosine Phosphorylation
- 3-Hydroxybenzoate:Coenzyme A Ligase from Cell Cultures of Centaurium erythraea: Isolation and Characterization
- Gel Chromatographic Characterization of the Hydrophobic Interaction of Glycosylphosphatidylinositol-Alkaline Phosphatase with Detergents
- Immunolabeling of CD3-Positive Lymphocytes with a Recombinant Single-Chain Antibody/Alkaline Phosphatase Conjugate
- Phage Display of RNase A and an Improved Method for Purification of Phages Displaying RNases
Articles in the same Issue
- The Twin-Arginine Translocation System: A Novel Means of Transporting Folded Proteins in Chloroplasts and Bacteria
- De Novo DNA Methylation at Nonrandom Founder Sites 5' from an Unmethylated Minimal Origin of DNA Replication in Latent Epstein-Barr Virus Genomes
- Trichostatin A Modulates Expression of p21waf1/cip1, Bcl-xL, ID1, ID2, ID3, CRAB2, GATA-2, hsp86 and TFIID/TAFII31 mRNA in Human Lung Adenocarcinoma Cells
- Effects of the Active Aldehyde Group Generated by RNA N-Glycosidase in the Sarcin/Ricin Domain of Rat 28S Ribosomal RNA on Peptide Elongation
- Responses to Peroxynitrite in Yeast: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) as a Sensitive Intracellular Target for Nitration and Enhancement of Chaperone Expression and Ubiquitination
- Enhancing the T R Transition of Insulin by Helix-Promoting Sequence Modifications at the N-Terminal B-Chain
- Structure-Dependent Effects of Glucose-Containing Analogs of Platelet Activating Factor (PAF) on Membrane Integrity
- Two Different Mechanisms for Activation of Cyclic PIP Synthase: by a G Protein or by Protein Tyrosine Phosphorylation
- 3-Hydroxybenzoate:Coenzyme A Ligase from Cell Cultures of Centaurium erythraea: Isolation and Characterization
- Gel Chromatographic Characterization of the Hydrophobic Interaction of Glycosylphosphatidylinositol-Alkaline Phosphatase with Detergents
- Immunolabeling of CD3-Positive Lymphocytes with a Recombinant Single-Chain Antibody/Alkaline Phosphatase Conjugate
- Phage Display of RNase A and an Improved Method for Purification of Phages Displaying RNases
