Table Of ContentCoverphotocredit:
Mulloy,B.,Rider,C.C.
TheBoneMorphogeneticProteinsandTheirAntagonists
VitaminsandHormones(2015)99,pp.63–90
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ROBERT S. HARRIS KENNETH V. THIMANN
Newton, Massachusetts University of California
Santa Cruz, California
JOHN A. LORRAINE
IRA G. WOOL
University of Edinburgh
Edinburgh, Scotland University of Chicago
Chicago, Illinois
PAUL L. MUNSON
EGON DICZFALUSY
University of North Carolina
Chapel Hill, North Carolina Karolinska Sjukhuset
Stockholm, Sweden
JOHN GLOVER
ROBERT OLSEN
University of Liverpool
Liverpool, England School of Medicine
State University of New York
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CONTRIBUTORS
PaulF.Austin
DepartmentofSurgery,DivisionofUrology,WashingtonUniversitySchoolofMedicine,
St.LouisChildren’sHospital,St.Louis,Missouri,USA
AnaClaudiaOliveiraCarreira
NUCEL-NETCEM(CellandMolecularTherapyCenter),InternalMedicineDepartment,
SchoolofMedicine,UniversityofSa˜oPaulo,Sa˜oPaulo,Brazil
SuvroChatterjee
VascularBiologyLab,AU-KBCResearchCentre,MITCampus,andDepartmentof
Biotechnology,AnnaUniversity,Chennai,India
JianQ.Feng
DepartmentofBiomedicalSciences,TexasA&MBaylorCollegeofDentistry,Dallas,Texas,
USA
RenatoAstorinoFilho
NUCEL-NETCEM(CellandMolecularTherapyCenter),InternalMedicineDepartment,
SchoolofMedicine,UniversityofSa˜oPaulo,Sa˜oPaulo,Brazil
Jose´ MauroGranjeiro
BioengineeringDivision,NationalInstituteofMetrology,Quality,andTechnology,Duque
deCaxias,andDepartmentofDentalMaterials,DentalSchool,FluminenseFederal
University,Niteroi,Brazil
JudithB.Grinspan
Children’sHospitalofPhiladelphia,andPerelmanSchoolofMedicine,Universityof
Pennsylvania,Philadelphia,Pennsylvania,USA
QiushaGuo
DepartmentofSurgery,DivisionofUrology,WashingtonUniversitySchoolofMedicine,
St.LouisChildren’sHospital,St.Louis,Missouri,USA
RobertJ.Hinton
DepartmentofBiomedicalSciences,TexasA&MBaylorCollegeofDentistry,Dallas,
Texas,USA
EijiroJimi
DivisionofMolecularSignalingandBiochemistry,DepartmentofHealthPromotion,
CenterforOralBiologicalResearch,KyushuDentalUniversity,Kitakyushu,Fukuoka,
Japan
JunjunJing
StateKeyLaboratoryofOralDiseases,WestChinaHospitalofStomatology,Sichuan
University,Chengdu,China
PiotrKraj
DepartmentofBiologicalSciences,OldDominionUniversity,Norfolk,Virginia,USA
xi
xii Contributors
MichalKuczma
CancerCenter,GeorgiaRegentsUniversity,Augusta,Georgia,USA
IsabelLaRosa
LaboratoryofAnimalBiotechnology,AgricultureFaculty,UniversityofBuenosAires
(UBA),BuenosAires,Argentina
ScottR.Manson
DepartmentofSurgery,DivisionofUrology,WashingtonUniversitySchoolofMedicine,
St.LouisChildren’sHospital,St.Louis,Missouri,USA
KatelynnH.Moore
DepartmentofSurgery,DivisionofUrology,WashingtonUniversitySchoolofMedicine,
St.LouisChildren’sHospital,St.Louis,Missouri,USA
ThomasD.Mueller
DepartmentPlantPhysiologyandBiophysics,Julius-von-SachsInstituteoftheUniversity
Wuerzburg,Wuerzburg,Germany
BarbaraMulloy
CentreforBiomedicalSciences,SchoolofBiologicalSciences,RoyalHolloway,University
ofLondon,Egham,Surrey,UnitedKingdom
SaranyaRajendran
VascularBiologyLab,AU-KBCResearchCentre,AnnaUniversity,MITCampus,
Chennai,India
ChrisC.Rider
CentreforBiomedicalSciences,SchoolofBiologicalSciences,RoyalHolloway,University
ofLondon,Egham,Surrey,UnitedKingdom
MarianaCorreaRossi
DepartmentofChemistryandBiochemistry,BiosciencesInstitute,UNESP,Universidade
EstadualPaulista,Botucatu,Brazil
JamilaH.Siamwala
DepartmentofOrthopaedicSurgery,UniversityofCalifornia,SanDiego,California,USA
MariCleideSogayar
NUCEL-NETCEM(CellandMolecularTherapyCenter),InternalMedicineDepartment,
SchoolofMedicine,UniversityofSa˜oPaulo,andChemistryInstitute,Biochemistry
Department,Sa˜oPaulo,Brazil
WillianFernandoZambuzzi
DepartmentofChemistryandBiochemistry,BiosciencesInstitute,UNESP,Universidade
EstadualPaulista,Botucatu,Brazil
PREFACE
Bone morphogenic (or morphogenetic) proteins (BMPs) represent a sub-
familyinthetransforminggrowthfactorbetasuperfamily.About20BMPs
are already known. First discovered in connection with their activities on
bone, they play key roles in bone formation and skeletal development
and in the differentiation of cartilage and chondrocytes. BMPs are consid-
eredaspotentialtreatmentsforbonehealingandforthelossofboneduring
spaceflight,especiallyinflightsofextendedduration.Additionally,itisnow
recognized that BMPs are involved in the development of several tissues
including limb buds, kidney, heart, eye, and skin. In kidney disease, for
example,BMPlevelsfall,openingthepossibilitythatBMPtreatmentmight
offer a beneficial therapeutic effect.
Thebasicinformation on the interactionof BMPs triggering activation
oftheirreceptorsaswellastheantagonistsofthisinteractionisknown.Fea-
turesofBMPsignalingalsoarenowbeingstudied.Thesignalingprocesses
involvenuclearfactorkappaBinsomecasesandalsoareknowntoaffectthe
process of myelination.
The chapters in this volume are arranged by first considering the basic
informationinthemechanismofBMPaction.Accordingly,thefirstchap-
ters deal with BMP–receptor interaction. T.D. Mueller describes the
“Mechanisms of BMP–Receptor Interaction and Activation.” In addition,
B. Mulloy and C.C. Rider report on “The Bone Morphogenetic Proteins
andTheirAntagonists.”Bothofthesechaptersdemonstratetheuseofthree-
dimensional crystal structures.
S.R.Manson,P.F.Austin,Q.Guo,andK.H.Moorereporton“BMP-7
Signaling and Its CriticalRoles in Kidney Development,theResponses to
RenalInjury,andChronicKidneyDisease.”E.Jimireportson“TheRole
of BMP Signaling and NF-κB Signaling on Osteoblastic Differentiation,
Cancer Development, and Vascular Diseases—Is the Activation of
NF-κB a Friend or Foe of BMP Function?” Additionally, in this vein,
M. Kuczma and P. Kraj review “Bone Morphogenic Protein Signaling
Regulates Development and Activation of CD4+ T Cells.” J. Grinspan
writes on “Bone Morphogenetic Proteins: Inhibitors of Myelination in
Development and Disease.” Continuing on the topic of development, I.
La Rosa describes “Bone Morphogenetic Proteins in Preimplantation
Embryos.”
xiii
xiv Preface
The following chapters refer to the effects on bone and cartilage.
J.H. Siamwala, S. Rajendran, and S. Chatterjee introduce “Strategies of
Manipulating BMP Signaling in Microgravity to Prevent Bone Loss.”
J. Jing, R.J. Hinton, andJ.Q. Feng review “Bmpr1aSignaling in Cartilage
Development and Endochondral Bone Formation.” The final chapter
covers “Bone Morphogenetic Proteins: Promising Molecules for Bone
Healing, Bioengineering, and Regenerative Medicine” authored by
A.C.O.Carreira,W.F.Zambuzzi,M.C.Rossi,R.A.Filho,M.C.Sogayar,
and J.M. Granjeiro.
TheillustrationonthecoverisFigure1ofChapter2byB.Mulloyand
C.C. Rider entitled “The Bone Morphogenetic Proteins and Their
Antagonists.”ItpresentsthecrystalstructureoftheBMPantagonistnoggin
complexed with the BMP-7 dimer (magenta) at the top. Noggin dimer is a
ribbonin redbelowwhileattheverybottom istheheparin-bindingsitein
yellow.
FinalprocessingofthisvolumewasfacilitatedbyHeleneKabes(Oxford,
UK) and Vignesh Tamilselvvan (Chennai, India).
GERALD LITWACK
North Hollywood, CA
June 17, 2015
CHAPTER ONE
–
Mechanisms of BMP Receptor
Interaction and Activation
Thomas D. Mueller1
DepartmentPlantPhysiologyandBiophysics,Julius-von-SachsInstituteoftheUniversityWuerzburg,
Wuerzburg,Germany
1Correspondingauthor:e-mailaddress:[email protected]
Contents
1. EvolutionaryExpansionandDiversificationoftheTransformingGrowthFactorβ
Superfamily 2
2. PhylogeneticAnalysisRevealsFourFunctionalSubfamiliesforTGFβLigands 5
3. ExpressionasProtease-ActivatedProproteinsandaCystine-KnotMotifinthe
C-TerminalMatureRegionasKeyFeaturesofTGFβLigandMembers 12
4. TGFβReceptorActivationandItsDownstreamSignalingCascade 18
5. TooFewReceptorsforTooManyLigandsLeadtoPromiscuity 22
6. MolecularMechanismstoEnsureLigand–ReceptorPromiscuityandSpecificity:
TheConceptofMultipleHotSpotsofBinding 25
7. MolecularMechanismstoEnsureLigand–ReceptorPromiscuityandSpecificity:
TheConceptofStructuralAdaptability 32
8. ConsequencesofPromiscuityandSpecificityintheTGFβSuperfamily:Conclusions 37
References 41
Abstract
Bone morphogenetic proteins (BMPs), together with the eponymous transforming
growthfactor(TGF)βandtheActivinsformtheTGFβsuperfamilyofligands.Thisprotein
familycomprisesmorethan30structurallyhighlyrelatedproteins,whichdeterminefor-
mation,maintenance,andregenerationoftissuesandorgans.Theirimportanceforthe
developmentofmulticellularorganismsisevidentfromtheirexistenceinallvertebrates
aswellasnonvertebrate animals.Fromtheirhighlyspecific functionsinvivoeithera
strict relation between a particular ligand and its cognate cellular receptor and/or a
stringentregulationtodefineadistincttemperospatialexpressionpatternforthevar-
iousligandsandreceptorisexpected.However,onlyalimitednumberofreceptorsare
foundtoservealargenumberofligandsthusimplicatinghighlypromiscuousligand–
receptor interactionsinstead. Sincein tissuesamultitude ofligandsare oftenfound,
whichsignalviaahighlyoverlappingsetofreceptors,thisraisesthequestionhowsuch
promiscuousinteractionsbetweendifferentligandsandtheirreceptorscangenerate
concertedandhighlyspecificcellularsignalsrequiredduringembryonicdevelopment
andtissuehomeostasis.
VitaminsandHormones,Volume99 #2015ElsevierInc. 1
ISSN0083-6729 Allrightsreserved.
http://dx.doi.org/10.1016/bs.vh.2015.06.003
2 ThomasD.Mueller
1. EVOLUTIONARY EXPANSION AND DIVERSIFICATION
OF THE TRANSFORMING GROWTH FACTOR β
SUPERFAMILY
Multicellularorganismsrequirecontinuousintercellularcommunica-
tionnotonlyduringtheirdevelopmentbutalsoforhomeostasisandsurvival.
Processes such as cell differentiation, proliferation, migration or apoptosis
dependonendocrine,paracrineorpossiblyautocrinestimuli,whichattheir
heartareoften,butnotexclusivelyexertedbyprotein–proteininteractions
at the cell surface involving a secreted (sometimes also membrane-
associated)growthfactor,andatransmembranereceptor.Duringevolution,
nature has “recycled” successful examples of above combinations thereby
forminglargerproteinfamilies,inwhichfurtherhomologousgrowthfactors
plustheirrespectivereceptorswereformedpossiblybygeneduplicationand
acquired additional functionalities necessary to cope with the increasing
complexity of the evolving organisms. The transforming growth factor β
(TGFβ)superfamilycomprisingTGFβs,Activins,andbonemorphogenetic
proteins (BMPs) as well as growth and differentiation factors (GDFs) pre-
sents a prime example of such a protein family with a few growth factors
in simple organisms like worms (five TGFβ ligands, for review: Savage-
Dunn,2005)andalargenumberofligandsinmammals(>30TGFβfactors
in human, for review: Feng & Derynck, 2005; Hinck, 2012; Mueller &
Nickel,2012;Fig.1A).AnevolutionaryexpansionintheTGFβsuperfamily
canbealsonotedfromtheobservationthathomologsofBMPs—incontrast
to senso strictu TGFβs and Activins—are already found in worms, whereas
homologsofActivinsappearforthefirsttimeinfliesandsensostrictuTGFβs
emergeinfishandamphibian(Newfeld,Wisotzkey,&Kumar,1999).This
suggests that BMPs are likely the founding members of this growth factor
family, which then diverged into Activins and TGFβ. Thus, TGFβs seem
to be the evolutionary youngest members despite serving as eponym of
the whole superfamily. The later emergence of Activins and TGFβs is also
consistent with their encoded functionalities. Activins modulate the
reproductiveaxis(Bilezikjian,Blount,Donaldson,&Vale,2006)andexert
regulatory roles in inflammation and immunity (for reviews: Aleman-
Muench & Soldevila, 2012; Hedger, Winnall, Phillips, & de Kretser,
2011),andTGFβsbeingimplicatedinthecontrolofimmunity(forreview:
Yoshimura & Muto, 2011) and wound healing (for review: Leask &
Abraham, 2004), functions that are not or differently implemented in
MechanismsofBMP–ReceptorInteractionandActivation 3
Figure1 (A)PhylogeneticanalysisoftheTGFβligandsuperfamily.TheTGFβscanbe
classifiedintofoursubgroupsindicatedontheleft:(I)sensustrictoTGFβs,(II)Activin/
Inhibins,(III)BMPs/GDFs,and(IV)others.TypeIandtypeIIreceptorrecruitmentisindi-
cated,theactivationofeithertheSMAD1/5/8orSMAD2/3pathwayismarkedbylightor
darkgray-shadedboxes,respectively.(B)PhylogeneticanalysisoftheTGFβreceptors
showingtheclassificationintotypeIandtypeIIreceptors.Lightanddarkgrayboxes
indicate the activation of either SMAD1/5/8 or SMAD2/3. (C) TGFβ proteins are
expressedaspre-proproteinscontainingasignalpeptide(SP),aprodomain,whichin
TGFβs is covalently dimerized by disulfide bonds (marked by asterisks), a proteolytic
processingsite(RXXR)andamatureregioncontainingthecharacteristiccystine-knot
motif comprisingsixconserved cysteine residues (marked bybars). Some TGFβs lack
aseventhcysteineresidue(markedbytwoasterisks)involvedincovalentdimerforma-
tion.(D)ArchitectureoftheTGFβreceptorscomprisingasignalpeptide(SP),anextra-
cellular ligand-binding domain (ECD), a single-span transmembrane element, and an
intracellularkinasedomain.TypeIreceptorsdifferbyanadditionalmembrane-proximal
glycine/serine-richmotif(GS-box).Furthermore,BMPRIIhasauniqueC-terminaldomain
(markedbyanasterisks),whichrecruitsadditionalsignalingproteins.
4 ThomasD.Mueller
simplerorganismssuchaswormsorinsects.ButnotonlyTGFβsandActivin
additionallyappearedlaterinevolution,butalsothenumberofBMPhomo-
logs expanded dramatically.
InCaenorhabditiselegans,fourofthefiveTGFβmembers,dbl1,daf7,tig2,
and tig3, could be mapped to the mammalian BMP orthologs, BMP5,
GDF8/11, BMP8, and BMP2 (for review: Gumienny & Savage-Dunn,
2013); however, the functional similarities seem limited. For instance,
dbl1 and daf7, which are involved in the regulation of body size in the
so-called Dauer larval development pathway, possibly exert a similar
growth-limiting function as found for GDF8/11 in vertebrates. Despite
theirlimitedhomologywithBMP8andBMP2,nofunctionshaveyetbeen
described for the C. elegans orthologs tig-2 and tig-3, but both members
might be involved in patterning. Unc129, whose mature region exhibits
limited sequence homology to mammalian BMP8 and GDF6, seems to
be involved in axon guidance and signals via a non-TGFβ related non-
canonical signaling pathway (Gumienny & Savage-Dunn, 2013). In flies,
seven TGFβ members have been identified of which the ligands dpp,
gbb, and screw can be mapped to the mammalian BMP2/4 and
BMP5/6/7 (Newfeld et al., 1999), myoglianin likely presents an ortholog
of GDF8/11 (Lo & Frasch, 1999), and dActivinβ, Dawdle and Maverick
are fly Activin-like ligands (Kutty et al., 1998; Nguyen, Parker, & Arora,
2000; Parker, Ellis, Nguyen, & Arora, 2006; Serpe & O’Connor, 2006).
Possibly due to the evolutionary smaller distance, the fly BMP orthologs
dpp, gbb, and screw exert in vivo function more closely related to their
vertebrate/mammalian counterparts. Dpp, the fly ortholog of BMP2 and
BMP4, is essential for correct dorsoventral patterning in fly (Irish &
Gelbart, 1987), a function it shares with BMP2/swirl in fish (Kishimoto,
Lee,Zon,Hammerschmidt,&Schulte-Merker,1997)andBMP4inmouse
(Winnier,Blessing,Labosky,&Hogan,1995).Drosophilagbbisinvolvedin
thedevelopmentofthefly’sintestinaltractortheeyessimilarlyasfoundfor
BMP6/7 in vertebrates (Helder et al., 1995; Luo et al., 1995; Perr, Ye, &
Gitelman, 1999; Wharton et al., 1999). On the contrary, the functions
encodedbydActivinβandthefurtherdistantActivin-likemembersDawdle
and Maverick seem to be more limited to neuronal morphogenesis com-
pared to their vertebrate homologs (Kutty et al., 1998; Nguyen et al.,
2000; Ting et al., 2007; Zhu et al., 2008).
Withtheemergenceofvertebrates,thenumberofTGFβmembersnot
only doubled as evident from the 14 and 19 TGFβ ligands in fish (two
Activin orthologs; Thisse, Wright, & Thisse, 2000) of Danio rerio are not
listedinMassague(2000)andamphibian(Xenopuslaevis),buttheirencoded
