Dataset Information

No evidence for vitamin K-dependent carboxylation of canine surfactant apoproteins, 28-36 kDa.

ABSTRACT: Recent research has shown that rat surfactant apoproteins (26-38 kDa) are vitamin K-dependent [Rannels, Gallaher, Wallin & Rannels (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 5952-5956]. We have investigated the effect of the vitamin K antagonist warfarin on this family of apoproteins in surfactant from dog lung. Our data suggest that warfarin does not interfere with synthesis and secretion of these proteins into dog lung surfactant. Abnormal surfactant apoproteins, produced in response to warfarin treatment of the dog, were also not found in lung surfactant. 4-Carboxyglutamic acid analysis of purified dog apoproteins also failed to detect the vitamin K-modification. When vitamin K-dependent 14C labelling of precursors of vitamin K-dependent proteins was carried out, fluorography of these precursors, when electrophoresed into SDS/polyacrylamide gels, revealed 14C-labelled proteins of apparent molecular mass 74, 46, 42, 34, 31 and 23 kDa. Antibodies produced against purified dog surfactant apoproteins recognized precursors of the surfactant apoproteins in lung microsomes but did not recognize any 14C-labelled carboxylase substrates. These precursors appeared on immunoblots with apparent molecular mass 29, 32, 33 and 50 kDa. Our data suggest that there are significant differences between this class of surfactant apoproteins in the rat and the dog.

SUBMITTER: Wallin R

PROVIDER: S-EPMC1149225 | biostudies-other | 1988 Jun

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:Periostin (PN, gene name POSTN) is an extracellular matrix protein that is up-regulated in bronchial epithelial cells and lung fibroblasts by TH-2 cytokines. Its paralog, TGF-?-induced protein (?ig-h3, gene name TGFBI), is also expressed in the lung and up-regulated in bronchial myofibroblasts by TGF-?. PN and ?ig-h3 contain fasciclin 1 modules that harbor putative recognition sequences for ?-glutamyl carboxylase and are annotated in UniProt as undergoing vitamin K-dependent ?-carboxylation of multiple glutamic acid residues. ?-carboxylation profoundly alters activities of other proteins subject to the modification, e.g., blood coagulation factors, and would be expected to alter the structure and function of PN and ?ig-h3. To analyze for the presence of ?-carboxylation, proteins extracted from fibrotic lung were reacted with monoclonal antibodies specific for PN, ?ig-h3, or modification with ?-carboxyglutamic acid (Gla). In Western blots of 1-dimensional gels, bands stained with anti-PN or -?ig-h3 did not match those stained with anti-Gla. In 2-dimensional gels, anti-PN-positive spots had pIs of 7.0 to >8, as expected for the unmodified protein, and there was no overlap between anti-PN-positive and anti-Gla-positive spots. Recombinant PN and blood coagulation factor VII were produced in HEK293 cells that had been transfected with vitamin K 2, 3-epoxide reductase C1 to optimize ?-carboxylation. Recombinant PN secreted from these cells did not react with anti-Gla antibody and had pIs similar to that found in extracts of fibrotic lung whereas secreted factor VII reacted strongly with anti-Gla antibody. Over 67% coverage of recombinant PN was achieved by mass spectrometry, including peptides with 19 of the 24 glutamates considered targets of ?-carboxylation, but analysis revealed no modification. Over 86% sequence coverage and three modified glutamic acid residues were identified in recombinant fVII. These data indicate that PN and ?ig-h3 are not subject to vitamin K-dependent ?-carboxylation.

Dataset Information

No evidence for vitamin K-dependent carboxylation of canine surfactant apoproteins, 28-36 kDa.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets