Identification of Isoforms of the Exocytosis-sensitive Phosphoprotein PP63/parafusin in Paramecium Tetraurelia and Demonstration of Phosphoglucomutase Activity

Overview

Journal Biochem J

Specialty Biochemistry

Date 1997 Apr 1

PMID 9173895

Citations 4

Authors

K Hauser

R Kissmehl

J Linder

J E Schultz

F Lottspeich

H Plattner

Affiliations

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Abstract

PP63 (parafusin) is a 63 kDa phosphoprotein which is very rapidly (within 80 ms) dephosphorylated (to P63) during triggered trichocyst exocytosis; this occurs selectively in exocytosis-competent Paramecium tetraurelia strains. In the present work, two cDNAs coding for PP63/parafusin have been isolated, one of which is a new isoform. These isoforms are 99.6% identical and are derived from two different genes. Similarity searches revealed 43-51% identity of the deduced amino acid sequences with known phosphoglucomutases from yeast and mammals. The sequences of two proteolytic peptides obtained from PP63/parafusin isolated from Paramecium are identical to parts of the amino acid sequence deduced from the major cDNA. The major cDNA was mutated from the macronuclear ciliate genetic code into the universal genetic code and expressed in Escherichia coli. The recombinant protein shows the same biochemical and immunological characteristics as the (P)P63/parafusin originally isolated from Paramecium. It has the same specific phosphoglucomutase activity as phosphoglucomutase from chicken muscle. We also show that recombinant P63-1 parafusin 1 is a substrate of an endogenous casein kinase from Paramecium, as is the originally isolated P63/parafusin. Polyclonal antibodies against recombinant P63-1/parafusin 1 were raised which recognized phosphoglucomutases from different sources. Thus we show that PP63/parafusin and phosphoglucomutase in Paramecium are identical.

Citing Articles

Multigene family encoding 3',5'-cyclic-GMP-dependent protein kinases in Paramecium tetraurelia cells.

Kissmehl R, Kruger T, Treptau T, Froissard M, Plattner H Eukaryot Cell. 2006; 5(1):77-91.

PMID: 16400170 PMC: 1360248. DOI: 10.1128/EC.5.1.77-91.2006.

Molecular identification of a calcium-inhibited catalytic subunit of casein kinase type 2 from Paramecium tetraurelia.

Vetter D, Kissmehl R, Treptau T, Hauser K, Kellermann J, Plattner H Eukaryot Cell. 2003; 2(6):1220-33.

PMID: 14665457 PMC: 326640. DOI: 10.1128/EC.2.6.1220-1233.2003.

Molecular characterization of a sarco(endo)plasmic reticulum Ca2+-ATPase gene from Paramecium tetraurelia and localization of its gene product to sub-plasmalemmal calcium stores.

Hauser K, Pavlovic N, Kissmehl R, Plattner H Biochem J. 1998; 334 ( Pt 1):31-8.

PMID: 9693098 PMC: 1219657. DOI: 10.1042/bj3340031.

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