Formulation	20mMTris,150mMNaCl,pH7.4
Storage	-80°C
Purity	>95%bySDS-PAGE
ActivityDetermination	N/A
ShelfLife(properlystored)	12months

AminoAcidSequenceofBovineOsteocalcin
Theaminoacidsequenceofbovineboneosteocalcin.Thepositionsoftheg-carboxyglutamicacidresiduesareindicatedbygamma.

SampleGelInformation:

Gel	Novex4-12%Bis-Tris
Load	Bovineosteocalcin,1µgperlane
Buffer	MES
Standard	SeeBluePlus2;Myosin(188kDa),PhosphorylaseB(98kDa),BSA(62kDa),GlutamicDehydrogenase(49kDa),AlcoholDehydrogenase(38kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(17kDa),Lysozyme(14kDa),Aprotinin(6kDa),Insulin,Bchain(3kDa).

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:

Localization	Bone,plasma
Modeofaction	Unknown
Molecularweight	5,800(1)
Extinctioncoefficient	E 1% 1cm,280nm =13.3(7)(calculated)
Isoelectricpoint	4.0-4.5(8)
Structure	Singlechain,oneintrachaindisulfidebond,Cys23-29.
Percentcarbohydrate	0%
Post-translationalmodifications	Threeglaresidues,17,21,24

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