Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs
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P. Gregory
Abstract
Despite its well characterised biochemistry, the physiological role of glycosylphosphatidylinositolspecific phospholipase D (GPIPLD) is unknown. Most of the previous studies investigating the distribution of GPI-PLD have focused on the human and bovine forms of the enzyme. Studies on mouse GPI-PLD are rare, partly due to the lack of a specific antimouse GPI-PLD antibody, but also due to the apparent low reactivity of existing antibodies to rodent GPI-PLDs. Here we describe the isolation of a mouse liver cDNA, the construction and expression of a recombinant enzyme and the generation of an affinitypurified rabbit antimouse GPI-PLD antiserum. The antibody shows good reactivity to partially purified murine and purified bovine GPI-PLD. In contrast, a rat antibovine GPI-PLD antibody shows no reactivity with the mouse enzyme and the two antibodies recognise different proteolytic fragments of the bovine enzyme. Comparison between the rodent, bovine and human enzymes indicates that small changes in the amino acid sequence of a short peptide in the mouse and bovine GPI-PLDs may contribute to the different reactivities of the two antisera. We discuss the implications of these results and stress the importance of antibody selection while investigating GPI-PLD in the mouse.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Metabolic Stability, Receptor Binding, cAMP Generation, Insulin Secretion and Antihyperglycaemic Activity of Novel N-Terminal Glu9-Substituted Analogues of Glucagon-Like Peptide-1
- Characterisation of Human Dipeptidyl Peptidase IV Expressed in Pichia pastoris. A Structural and Mechanistic Comparison between the Recombinant Human and the Purified Porcine Enzyme
- Effects of Iron Limitation on the Respiratory Chain and the Membrane Cytochrome Pattern of the Euryarchaeon Halobacterium salinarum
- Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs
- Substrate Specificity of Glutaminyl Cyclases from Plants and Animals
- The Binding of Haptoglobin to Apolipoprotein AI: Influence of Hemoglobin and Concanavalin A
- A Novel Water-Soluble and Cell-Permeable Calpain Inhibitor Protects Myocardial and Mitochondrial Function in Postischemic Reperfusion
- Potent Bivalent Inhibition of Human Tryptase-? by a Synthetic Inhibitor
- Aza-Peptide Epoxides: Potent and Selective Inhibitors of Schistosoma mansoni and Pig Kidney Legumains (Asparaginyl Endopeptidases)
- Semisynthesis and Application of Carboxyfluorescein-Labelled Biologically Active Human Interleukin-8
- Acknowledgment
- Content Index
- Author Index
- Subject Index
Articles in the same Issue
- Metabolic Stability, Receptor Binding, cAMP Generation, Insulin Secretion and Antihyperglycaemic Activity of Novel N-Terminal Glu9-Substituted Analogues of Glucagon-Like Peptide-1
- Characterisation of Human Dipeptidyl Peptidase IV Expressed in Pichia pastoris. A Structural and Mechanistic Comparison between the Recombinant Human and the Purified Porcine Enzyme
- Effects of Iron Limitation on the Respiratory Chain and the Membrane Cytochrome Pattern of the Euryarchaeon Halobacterium salinarum
- Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs
- Substrate Specificity of Glutaminyl Cyclases from Plants and Animals
- The Binding of Haptoglobin to Apolipoprotein AI: Influence of Hemoglobin and Concanavalin A
- A Novel Water-Soluble and Cell-Permeable Calpain Inhibitor Protects Myocardial and Mitochondrial Function in Postischemic Reperfusion
- Potent Bivalent Inhibition of Human Tryptase-? by a Synthetic Inhibitor
- Aza-Peptide Epoxides: Potent and Selective Inhibitors of Schistosoma mansoni and Pig Kidney Legumains (Asparaginyl Endopeptidases)
- Semisynthesis and Application of Carboxyfluorescein-Labelled Biologically Active Human Interleukin-8
- Acknowledgment
- Content Index
- Author Index
- Subject Index
