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

DomainStructureofHumanOsteonectin
Thedomainstructureofhumanosteonectinisillustrated,where:CHO=potentialglycosylationsites,OP=potentialserinephophorylationsites,EF-Hand=regionshomologoustocalciumbindingdomainsincalmodulin,andacidic=NH2-terminalacidicdomain.

SampleGelInformation:

Gel	Novex4-12%Bis-Tris
Load	Humanosteonectin,1µgperlane
Buffer	MOPS
Standard	SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa)

Overview:

Osteonectinisanacidic,noncollagenousglycoprotein(Mr=29,000)originallyisolatedfromfetalandadultbovinebonematrix(1-5).InvitrobovineboneosteonectinbindstypeIcollagen,calcium(Kd=3x10-7M)andhydroxyapatite(Kd=8x10-8M)andhasbeenshowntobeapotentinhibitorofhydroxyapatiteseededcrystalgrowth(6).Inthiscontextithasbeensuggestedthatosteonectinmayplayanimportantroleintheregulationofbonemetabolismbybindinghydroxyapatitetocollagen.Recently,proteinshomologoustoosteonectinhavebeenidentifiedinanumberofcelltypes,mostofwhichareassociatedwithextracellularmatrixproduction(7).Theaminoacidsequence(fromCDNAsequences)ofoneoftheseproteins,humanplacentalSPARCisidenticaltohumanboneosteonectin.

Osteonectinhasalsobeenidentifiedasanalphagranulecomponentofhumanplateletsandissecretedduringactivation.Asmallportionofthesecretedosteonectinisexpressedontheplateletcellsurfaceinanactivationdependentmanner(8).Purifiedplateletosteonectinisasinglechainmoleculewhichexhibitsaslightlylargerapparentmolecularweightthanthatofosteonectinderivedfrombone(8).TheNH2-terminalsequencesofplateletandbone-derivedosteonectinareidentical,butthetwoproteinsdifferwithregardtotheextentotglycosylation(8-10).

HumanplateletosteonectinisisolatedfromthrombinactivatedplateletsasdescribedbyKelm,etal.(8).BovineboneosteonectinisisolatedfromdemineralizedbonebythemethodofRombergetal.(2).Bothproteinsaresuppliedin0.02MTris,0.15MNaCl,pH7.4,andshouldbestoredat-80oC.PurityisjudgedbySDS-PAGEanalysis.

Properties:

Localization	Bone,platelets,plasma(0.9ug/ml)*(5),serum(2.6ug/ml)**(5).
Modeofaction	Unknown
Molecularweight	32,700(humanbone,fromcDNA)(1) 29,000(bovinebone,bysedimentationequilibrium)(3)
Extinctioncoefficient	E 1% 1cm,280nm =8.0
Isoelectricpoint	5.5***(3)
Structure	Singlechain,NH2terminalacidicdomain,cysteinerichserpinhomologydomain,2EFhanddomains.
Post-translationalmodifications	Phosphoserine(11)
Notes	*DeterminedforHumanPlasma **DeterminedforHumanSerum ***DeterminedforBovineBone

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