SampleGelInformation:

Overview:

Osteocalcin,alsoknownasBone-Gla-Protein(BGP),isasinglechain(Mr=5800)vitaminKdependentproteinproducedbyosteoblastsandfoundinhighconcentrationsinbone(1-4).Post-translationalmodificationbyavitaminKdependentcarboxylaseproducesthreeg-carboxyglutamicacidresiduesatpositions17,21and24.Thematureproteincontains49aminoacidsandasingleintrachaindisulfidebondjoiningCys23toCys29.Thesecondarystructureishighlycalciumdependentandcontains14%α-helix,20%β-sheetand67%randomforminthepresenceofcalcium,and1%α-helix,20%β-sheetand79%randomformintheabsenceofcalcium(4).

BGPbindstophospholipidvesiclesinthepresenceofcalciumions(Kd=6x10-6M)(5).BGPalsobindshydroxyapatiteandisanefficientinhibitorofhydroxyapatiteseededcrystalgrowth,suggestingaregulatoryroleinbonemineralization(6).AlthoughthefundamentalroleofBGPinvivoremainselusive,invitroexperimentationsuggestsrolesinbothboneresorptionandboneformation.BGPhasbeenshowntofunctionasachemo-attractantinaninvitromacrophagemediatedboneresorptionsystem.TheserumconcentrationofBGPhasbeenlinkedtoboneformationrates,andBGPlevelsarecurrentlyusedinresearchassessmentofosteoporosisandothermetabolicbonediseasestates(7).

OsteocalcinispreparedfrombovinebonegenerallyasdescribedbyPriceetal.(1).ThisinvolvesdemineralizationinEDTA,sizeexclusionandionexchangechromatography.Thepurifiedproteinissupplied50%(vol/vol)glycerol/0.01MTris,0.075MNaCl,pH7.4,andshouldbestoredat-20°C.Humanosteocalcinispreparedfromextractsofacid-demineralizedbone,usingimmunoaffinitychromatography.Thehumanproteinissuppliedin0.02MTris,0.15MNaCl,2mMCaCl2,pH7.4andshouldbestoredat-80°C.PurityisdeterminedbySDS-PAGE.

Properties:

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