Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2

Petra Seemann; Raphaela Schwappacher; Klaus Wilbrandt Kjær; Deborah Krakow; Katarina Lehmann; Katherine Dawson; Sigmar Stricker; Jens Pohl; Frank Plöger; Eike Staub; Joachim Nickel; Walter Sebald; Petra Knaus; Stefan Mundlos

doi:10.1172/JCI25118

Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2

Petra Seemann, Raphaela Schwappacher, Klaus Wilbrandt Kjær, Deborah Krakow, Katarina Lehmann, Katherine Dawson, Sigmar Stricker, Jens Pohl, Frank Plöger, Eike Staub, Joachim Nickel, Walter Sebald, Petra Knaus, Stefan Mundlos

Department of Cellular and Molecular Medicine

154 Citations (Scopus)

Abstract

Here we describe 2 mutations in growth and differentiation factor 5 (GDF5) that alter receptor-binding affinities. They cause brachydactyly type A2 (L441P) and symphalangism (R438L), conditions previously associated with mutations in the GDF5 receptor bone morphogenetic protein receptor type 1b (BMPR1B) and the BMP antagonist NOGGIN, respectively. We expressed the mutant proteins in limb bud micromass culture and treated ATDC5 and C2C12 cells with recombinant GDF5. Our results indicated that the L441P mutant is almost inactive. The R438L mutant, in contrast, showed increased biological activity when compared with WT GDF5. Biosensor interaction analyses revealed loss of binding to BMPR1A and BMPR1B ectodomains for the L441P mutant, whereas the R438L mutant showed normal binding to BMPR1B but increased binding to BMPR1A, the receptor normally activated by BMP2. The binding to NOGGIN was normal for both mutants. Thus, the brachydactyly type A2 phenotype (L441P) is caused by inhibition of the ligand-receptor interaction, whereas the symphalangism phenotype (R438L) is caused by a loss of receptor-binding specificity, resulting in a gain of function by the acquisition of BMP2-like properties. The presented experiments have identified some of the main determinants of GDF5 receptor-binding specificity in vivo and open new prospects for generating antagonists and superagonists of GDF5.

Original language	English
Journal	Journal of Clinical Investigation
Volume	115
Issue number	9
Pages (from-to)	2373-81
Number of pages	9
ISSN	0021-9738
DOIs	https://doi.org/10.1172/JCI25118
Publication status	Published - 2005

Keywords

Amino Acid Sequence
Animals
Bone Morphogenetic Protein Receptors, Type I
Bone Morphogenetic Proteins
Carrier Proteins
Cell Differentiation
Cell Line
Embryonic Structures
Fingers
Growth Differentiation Factor 5
Humans
In Situ Hybridization
Limb Deformities, Congenital
Mice
Molecular Sequence Data
Phenotype
Point Mutation
Protein Binding
Protein Conformation
Recombinant Proteins
Sequence Alignment
Tissue Culture Techniques

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10.1172/JCI25118

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Seemann, P., Schwappacher, R., Kjær, K. W., Krakow, D., Lehmann, K., Dawson, K., Stricker, S., Pohl, J., Plöger, F., Staub, E., Nickel, J., Sebald, W., Knaus, P., & Mundlos, S. (2005). Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2. Journal of Clinical Investigation, 115(9), 2373-81. https://doi.org/10.1172/JCI25118

Seemann, P, Schwappacher, R, Kjær, KW, Krakow, D, Lehmann, K, Dawson, K, Stricker, S, Pohl, J, Plöger, F, Staub, E, Nickel, J, Sebald, W, Knaus, P & Mundlos, S 2005, 'Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2', Journal of Clinical Investigation, vol. 115, no. 9, pp. 2373-81. https://doi.org/10.1172/JCI25118

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title = "Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2",

abstract = "Here we describe 2 mutations in growth and differentiation factor 5 (GDF5) that alter receptor-binding affinities. They cause brachydactyly type A2 (L441P) and symphalangism (R438L), conditions previously associated with mutations in the GDF5 receptor bone morphogenetic protein receptor type 1b (BMPR1B) and the BMP antagonist NOGGIN, respectively. We expressed the mutant proteins in limb bud micromass culture and treated ATDC5 and C2C12 cells with recombinant GDF5. Our results indicated that the L441P mutant is almost inactive. The R438L mutant, in contrast, showed increased biological activity when compared with WT GDF5. Biosensor interaction analyses revealed loss of binding to BMPR1A and BMPR1B ectodomains for the L441P mutant, whereas the R438L mutant showed normal binding to BMPR1B but increased binding to BMPR1A, the receptor normally activated by BMP2. The binding to NOGGIN was normal for both mutants. Thus, the brachydactyly type A2 phenotype (L441P) is caused by inhibition of the ligand-receptor interaction, whereas the symphalangism phenotype (R438L) is caused by a loss of receptor-binding specificity, resulting in a gain of function by the acquisition of BMP2-like properties. The presented experiments have identified some of the main determinants of GDF5 receptor-binding specificity in vivo and open new prospects for generating antagonists and superagonists of GDF5.",

keywords = "Amino Acid Sequence, Animals, Bone Morphogenetic Protein Receptors, Type I, Bone Morphogenetic Proteins, Carrier Proteins, Cell Differentiation, Cell Line, Embryonic Structures, Fingers, Growth Differentiation Factor 5, Humans, In Situ Hybridization, Limb Deformities, Congenital, Mice, Molecular Sequence Data, Phenotype, Point Mutation, Protein Binding, Protein Conformation, Recombinant Proteins, Sequence Alignment, Tissue Culture Techniques",

author = "Petra Seemann and Raphaela Schwappacher and Kj{\ae}r, {Klaus Wilbrandt} and Deborah Krakow and Katarina Lehmann and Katherine Dawson and Sigmar Stricker and Jens Pohl and Frank Pl{\"o}ger and Eike Staub and Joachim Nickel and Walter Sebald and Petra Knaus and Stefan Mundlos",

year = "2005",

doi = "10.1172/JCI25118",

language = "English",

volume = "115",

pages = "2373--81",

journal = "Journal of Clinical Investigation",

issn = "0021-9738",

publisher = "American Society for Clinical Investigation",

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TY - JOUR

T1 - Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2

AU - Seemann, Petra

AU - Schwappacher, Raphaela

AU - Kjær, Klaus Wilbrandt

AU - Krakow, Deborah

AU - Lehmann, Katarina

AU - Dawson, Katherine

AU - Stricker, Sigmar

AU - Pohl, Jens

AU - Plöger, Frank

AU - Staub, Eike

AU - Nickel, Joachim

AU - Sebald, Walter

AU - Knaus, Petra

AU - Mundlos, Stefan

PY - 2005

Y1 - 2005

N2 - Here we describe 2 mutations in growth and differentiation factor 5 (GDF5) that alter receptor-binding affinities. They cause brachydactyly type A2 (L441P) and symphalangism (R438L), conditions previously associated with mutations in the GDF5 receptor bone morphogenetic protein receptor type 1b (BMPR1B) and the BMP antagonist NOGGIN, respectively. We expressed the mutant proteins in limb bud micromass culture and treated ATDC5 and C2C12 cells with recombinant GDF5. Our results indicated that the L441P mutant is almost inactive. The R438L mutant, in contrast, showed increased biological activity when compared with WT GDF5. Biosensor interaction analyses revealed loss of binding to BMPR1A and BMPR1B ectodomains for the L441P mutant, whereas the R438L mutant showed normal binding to BMPR1B but increased binding to BMPR1A, the receptor normally activated by BMP2. The binding to NOGGIN was normal for both mutants. Thus, the brachydactyly type A2 phenotype (L441P) is caused by inhibition of the ligand-receptor interaction, whereas the symphalangism phenotype (R438L) is caused by a loss of receptor-binding specificity, resulting in a gain of function by the acquisition of BMP2-like properties. The presented experiments have identified some of the main determinants of GDF5 receptor-binding specificity in vivo and open new prospects for generating antagonists and superagonists of GDF5.

AB - Here we describe 2 mutations in growth and differentiation factor 5 (GDF5) that alter receptor-binding affinities. They cause brachydactyly type A2 (L441P) and symphalangism (R438L), conditions previously associated with mutations in the GDF5 receptor bone morphogenetic protein receptor type 1b (BMPR1B) and the BMP antagonist NOGGIN, respectively. We expressed the mutant proteins in limb bud micromass culture and treated ATDC5 and C2C12 cells with recombinant GDF5. Our results indicated that the L441P mutant is almost inactive. The R438L mutant, in contrast, showed increased biological activity when compared with WT GDF5. Biosensor interaction analyses revealed loss of binding to BMPR1A and BMPR1B ectodomains for the L441P mutant, whereas the R438L mutant showed normal binding to BMPR1B but increased binding to BMPR1A, the receptor normally activated by BMP2. The binding to NOGGIN was normal for both mutants. Thus, the brachydactyly type A2 phenotype (L441P) is caused by inhibition of the ligand-receptor interaction, whereas the symphalangism phenotype (R438L) is caused by a loss of receptor-binding specificity, resulting in a gain of function by the acquisition of BMP2-like properties. The presented experiments have identified some of the main determinants of GDF5 receptor-binding specificity in vivo and open new prospects for generating antagonists and superagonists of GDF5.

KW - Amino Acid Sequence

KW - Animals

KW - Bone Morphogenetic Protein Receptors, Type I

KW - Bone Morphogenetic Proteins

KW - Carrier Proteins

KW - Cell Differentiation

KW - Cell Line

KW - Embryonic Structures

KW - Fingers

KW - Growth Differentiation Factor 5

KW - Humans

KW - In Situ Hybridization

KW - Limb Deformities, Congenital

KW - Mice

KW - Molecular Sequence Data

KW - Phenotype

KW - Point Mutation

KW - Protein Binding

KW - Protein Conformation

KW - Recombinant Proteins

KW - Sequence Alignment

KW - Tissue Culture Techniques

U2 - 10.1172/JCI25118

DO - 10.1172/JCI25118

M3 - Journal article

C2 - 16127465

SN - 0021-9738

VL - 115

SP - 2373

EP - 2381

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

IS - 9

ER -

Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2

Abstract

Keywords

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