Specificity and Structure of a High Affinity Activin Receptor-like Kinase 1 (ALK1) Signaling Complex

Townson, Sharon A.; Martı́nez-Hackert, Erik; Greppi, Chloe; Lowden, Patricia; Sako, Dianne; Liu, June; Ucran, Jeffrey A.; Liharska, Katia; Underwood, Kathryn; Seehra, Jasbir; Kumar, Ravindra; Grinberg, Asya V.

doi:10.1074/jbc.m112.377960

Cited by 156 publications

(237 citation statements)

References 37 publications

(44 reference statements)

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“…4F). The EC 50 values for type I and II receptors agree with previous affinity measurements with BMP9 (16,17). Interestingly, the BMP7 prodomain also competes binding of BMP7 to type II but not type I receptors (18), consistent with the open-armed conformation of pro-BMP7 (Fig.…”

Section: Resultssupporting

confidence: 79%

“…Furthermore, ALK1, which primarily functions as a receptor for BMP9 and BMP10, has recently been implicated as an important target of antiangiogenic tumor therapy (17). Together with the finding that BMP9 complexed with its prodomain circulates in the bloodstream at physiologically relevant concentrations (11), our finding that the BMP9 prodomain blocks binding to ActRIIA and alters selectivity for ActRIIB compared with BMPRII has important implications for BMP9 signaling in vivo.…”

Section: Resultsmentioning

confidence: 55%

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Structure of bone morphogenetic protein 9 procomplex

Brown

Gao

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

104

218

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Bone morphogenetic proteins (BMPs) belong to the TGF-β family, whose 33 members regulate multiple aspects of morphogenesis. TGF-β family members are secreted as procomplexes containing a small growth factor dimer associated with two larger prodomains. As isolated procomplexes, some members are latent, whereas most are active; what determines these differences is unknown. Here, studies on pro-BMP structures and binding to receptors lead to insights into mechanisms that regulate latency in the TGF-β family and into the functions of their highly divergent prodomains. The observed open-armed, nonlatent conformation of pro-BMP9 and pro-BMP7 contrasts with the cross-armed, latent conformation of pro-TGF-β1. Despite markedly different arm orientations in pro-BMP and pro-TGF-β, the arm domain of the prodomain can similarly associate with the growth factor, whereas prodomain elements N-and C-terminal to the arm associate differently with the growth factor and may compete with one another to regulate latency and stepwise displacement by type I and II receptors. Sequence conservation suggests that pro-BMP9 can adopt both crossarmed and open-armed conformations. We propose that interactors in the matrix stabilize a cross-armed pro-BMP conformation and regulate transition between cross-armed, latent and open-armed, nonlatent pro-BMP conformations.

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Section: Resultssupporting

confidence: 79%

Section: Resultsmentioning

confidence: 55%

Structure of bone morphogenetic protein 9 procomplex

Brown

Gao

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

104

218

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“…This approach of removing murine content through mutations of tolerated germline residues is possible without the knowledge of the antibody:antigen complex crystal structure. This can be accomplished through experimental screening and selection methods like those described in Townson et al 57 There they described a method of phage display using germline substitutions in a binary selection library. From their CDR germlining of 3 different antibodies they were able to remove 32 to 53 % of the non-human residues (11 to 16 total residues) from the CDRs H1, H2, L1, L2 and L3 left after CDR grafting.…”

Section: Discussionmentioning

confidence: 99%

“…52,53 Of these, there are several available complex crystal structures showing a conserved binding mode of ActRIIB binding to the fingers region of activin A, 54,55 BMP-2, 56 and BMP-9. 57 Analysis of available structural data showing the conserved binding modes to ActRIIB, the sequence homology of the predicted binding interface of these homologs to myostatin, similar binding affinities, and correlated changes of affinity associated with ActRIIB mutations predicted to be in the binding interface, suggests that myostatin should bind to ActRIIB with the same binding mode as these other TGF-b family members (See Supplementary Results and Figs. S3 and S4).…”

Section: Epitope Overlapmentioning

confidence: 99%

Beyond CDR-grafting: Structure-guided humanization of framework and CDR regions of an anti-myostatin antibody

Apgar

Mader

Agostinelli

et al. 2016

mAbs

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Antibodies are an important class of biotherapeutics that offer specificity to their antigen, long half-life, effector function interaction and good manufacturability. The immunogenicity of non-human-derived antibodies, which can be a major limitation to development, has been partially overcome by humanization through complementarity-determining region (CDR) grafting onto human acceptor frameworks. The retention of foreign content in the CDR regions, however, is still a potential immunogenic liability. Here, we describe the humanization of an anti-myostatin antibody utilizing a 2-step process of traditional CDR-grafting onto a human acceptor framework, followed by a structure-guided approach to further reduce the murine content of CDR-grafted antibodies. To accomplish this, we solved the co-crystal structures of myostatin with the chimeric (Protein Databank (PDB) id 5F3B) and CDR-grafted antimyostatin antibody (PDB id 5F3H), allowing us to computationally predict the structurally important CDR residues as well as those making significant contacts with the antigen. Structure-based rational design enabled further germlining of the CDR-grafted antibody, reducing the murine content of the antibody without affecting antigen binding. The overall "humanness" was increased for both the light and heavy chain variable regions.

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“…Activin receptor-like kinase-1 (ALK1), the ACVRL1 gene product, is a type I receptor in the transforming growth factor-b (TGFb) superfamily with high affinity for BMP9 and BMP10 (11). Multiple lines of evidence implicate the BMP9/BMP10/ALK1 pathway as a key regulator of angiogenesis.…”

Section: Introductionmentioning

confidence: 99%