Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs

Verfasser / Beitragende:
[P. Gregory, A. Ziemiecki, G. Zürcher, U. Brodbeck, P. Bütikofer]
Ort, Verlag, Jahr:
2003
Enthalten in:
Biological Chemistry, 384/12(2003-12-25), 1575-1582
Format:
Artikel (online)
ID: 378856014
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245 0 0 |a Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs  |h [Elektronische Daten]  |c [P. Gregory, A. Ziemiecki, G. Zürcher, U. Brodbeck, P. Bütikofer] 
520 3 |a 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. 
540 |a Copyright © 2003 by Walter de Gruyter GmbH & Co. KG 
690 7 |a Biochemistry  |2 nationallicence 
690 7 |a Molecular biology  |2 nationallicence 
690 7 |a Cellular biology  |2 nationallicence 
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700 1 |a Ziemiecki  |D A.  |4 aut 
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700 1 |a Brodbeck  |D U.  |4 aut 
700 1 |a Bütikofer  |D P.  |4 aut 
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