Periostin, a member of a novel family of vitamin K-dependent proteins, is expressed by mesenchymal stromal cells.
The modification of glutamic acid residues to gamma-carboxyglutamic acid (Gla) is a post-translational modification catalyzed by the vitamin K-dependent enzyme gamma-glutamylcarboxylase. Despite ubiquitous expression of the gamma-carboxylation machinery in mammalian tissues, only 12 Gla-containing proteins have so far been identified in humans. Because bone tissue is the second most abundant source of Gla-containing proteins after the liver, we sought to identify Gla proteins secreted by bone marrow-derived mesenchymal stromal cells (MSCs). We used a proteomics approach to screen the secretome of MSCs with a combination of two-dimensional gel electrophoresis and tandem mass spectrometry. The most abundant Gla-containing protein secreted by MSCs was identified as periostin, a previously unrecognized gamma-carboxylated protein. In silico amino acid sequence analysis of periostin demonstrated the presence of four consensus gamma-carboxylase recognition sites embedded within fasciclin-like protein domains. The carboxylation of periostin was confirmed by immunoprecipitation and purification of the recombinant protein. Carboxylation of periostin could be inhibited by warfarin in MSCs, demonstrating its dependence on the presence of vitamin K. We were able to demonstrate localization of carboxylated periostin to bone nodules formed by MSCs in vitro, suggesting a role in extracellular matrix mineralization. Our data also show that another fasciclin I-like protein, betaig-h3, contains Gla. In conclusion, periostin is a member of a novel vitamin K-dependent gamma-carboxylated protein family characterized by the presence of fasciclin domains. Furthermore, carboxylated periostin is produced by bone-derived cells of mesenchymal lineage and is abundantly found in mineralized bone nodules in vitro.
Coutu
Daniel L
DL
Division of Experimental Medicine, McGill University, Montreal, Quebec, Canada.
Wu
Jian Hui
JH
Monette
Anne
A
Rivard
Georges-Etienne
GE
Blostein
Mark D
MD
Galipeau
Jacques
J
D000595:Amino Acid Sequence; D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation; D005786:Gene Expression Regulation; D006801:Humans; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D017154:Stromal Cells / Q000166:cytology* / Q000378:metabolism; D014812:Vitamin K / Q000737:chemistry*
D015815:Cell Adhesion Molecules; C512835:POSTN protein, human; D014812:Vitamin K