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First published online 16 December 2003
doi: 10.1242/jcs.00909

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The role of annexin 2 in osteoblastic mineralization

Jennifer M. Gillette¹ and Sheila M. Nielsen-Preiss^2,*,

¹ Department of Cellular and Developmental Biology, University of Colorado Health Sciences Center, Denver, CO 80262, USA
² Department of Orthopaedics, University of Colorado Health Sciences Center, Denver, CO 80262, USA

Author for correspondence (e-mail: sheila.preiss{at}uchsc.edu)

Accepted 17 September 2003

While the basic cellular contributions to bone differentiation and mineralization are widely accepted, the regulation of these processes at the intracellular level remains inadequately understood. Our laboratory recently identified annexin 2 as a protein involved in osteoblastic mineralization. Annexin 2 was overexpressed twofold in SaOSLM2 osteoblastic cells as a fusion protein with green fluorescent protein. The overexpression of annexin 2 led to an increase in alkaline phosphatase activity as well as an increase in mineralization. Our data suggest that the increase in alkaline phosphatase activity does not result from increased alkaline phosphatase transcript or protein levels; therefore we evaluated mechanism of action. We determined that both annexin 2 and alkaline phosphatase activity were localized to membrane microdomains called lipid rafts in osteoblastic cells. Annexin 2 overexpression resulted in an increase in alkaline phosphatase activity that was associated with lipid microdomains in a cholesterol-dependent manner. Furthermore, disruption of lipid rafts with a cholesterol sequestering agent or reduction of annexin 2 expression by specific antisense oligonucleotides each resulted in diminished mineralization. Therefore, intact lipid rafts containing annexin 2 appear to be important for alkaline phosphatase activity and may facilitate the osteoblastic mineralization process.

Key words: Lipid rafts, Bone, Alkaline phosphatase, Annexin 2, Osteoblast

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