Asymmetric Mode of Ca2+-S100A4 Interaction with Nonmuscle Myosin IIA Generates Nanomolar Affinity Required for Filament Remodeling

Elliott, P.R.; Irvine, Andrew J; Jung, Hyun Suk; Tozawa, Keiichi; Pastok, Martyna W.; Picone, Remigio; Badyal, Sandip K.; Basran, Jaswir; Rudland, Philip S.; Barraclough, Roger; Lian, Lu‐Yun; Bagshaw, Clive R.; Kriajevska, Marina; Barsukov, Igor

doi:10.1016/j.str.2012.02.002

Cited by 53 publications

(94 citation statements)

References 42 publications

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“…While the present manuscript was under revision, we became aware that Elliott et al (52) determined the NMR structure of wild-type S100A4 in complex with an NMIIA fragment and reached very similar conclusions. Fig.…”

Section: Methodssupporting

confidence: 54%

Crystal structure of the S100A4–nonmuscle myosin IIA tail fragment complex reveals an asymmetric target binding mechanism

Kiss

Duelli

Radnai

et al. 2012

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

120

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S100A4 is a member of the S100 family of calcium-binding proteins that is directly involved in tumor metastasis. It binds to the nonmuscle myosin IIA (NMIIA) tail near the assembly competence domain (ACD) promoting filament disassembly, which could be associated with increasing metastatic potential of tumor cells. Here, we investigate the mechanism of S100A4-NMIIA interaction based on binding studies and the crystal structure of S100A4 in complex with a 45-residue-long myosin heavy chain fragment. Interestingly, we also find that S100A4 binds as strongly to a homologous heavy chain fragment of nonmuscle myosin IIC as to NMIIA. The structure of the S100A4-NMIIA complex reveals a unique mode of interaction in the S100 family: A single, predominantly α-helical myosin chain is wrapped around the Ca 2þ -bound S100A4 dimer occupying both hydrophobic binding pockets. Thermal denaturation experiments of coiled-coil forming NMIIA fragments indicate that the coiled-coil partially unwinds upon S100A4 binding. Based on these results, we propose a model for NMIIA filament disassembly: Part of the random coil tailpiece and the C-terminal residues of the coiled-coil are wrapped around an S100A4 dimer disrupting the ACD and resulting in filament dissociation. The description of the complex will facilitate the design of specific drugs that interfere with the S100A4-NMIIA interaction.

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Section: Methodssupporting

confidence: 54%

Crystal structure of the S100A4–nonmuscle myosin IIA tail fragment complex reveals an asymmetric target binding mechanism

Kiss

Duelli

Radnai

et al. 2012

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

120

View full text Add to dashboard Cite

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“…Additionally, we found that NMIIC binds to S100A4 with similar affinity than NMIIA despite the two isoforms having rather different disordered tailpieces [20]. Finally, it is the disordered segment of the NMIIA peptide that has the most diverse conformation in existing S100A4-NMIIA complex structures further supporting the assumption that this element is not a key determinant of the interaction [20][21][22].…”

Section: Introductionsupporting

confidence: 61%

Structural determinants governing S100A4‐induced isoform‐selective disassembly of nonmuscle myosin II filaments

et al. 2016

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The Ca 2+ -binding protein S100A4 interacts with the C terminus of nonmuscle myosin IIA (NMIIA) causing filament disassembly, which is correlated with an increased metastatic potential of tumor cells. Despite high sequence similarity of the three NMII isoforms, S100A4 discriminates against binding to NMIIB. We searched for structural determinants of this selectivity. Based on paralog scanning using phage display, we identified a single position as major determinant of isoform selectivity. Reciprocal single amino acid replacements showed that at position 1907 (NMIIA numbering), the NMIIA/NMIIC-specific alanine provides about 60-fold higher affinity than the NMIIB-specific asparagine. The structural background of this can be explained in part by a communication between the two consecutive a-helical binding segments. This communication is completely abolished by the Ala-to-Asn substitution. Mutual swapping of the disordered tailpieces only slightly affects the affinity of the NMII chimeras. Interestingly, we found that the tailpiece and position 1907 act in a nonadditive fashion. Finally, we also found that the higher stability of the C-terminal coiled-coil region of NMIIB also discriminates against interaction with S100A4. Our results clearly show that the isoform-selective binding of S100A4 is determined at multiple levels in the structure of the three NMII isoforms and the corresponding functional elements of NMII act synergistically with one another resulting in a complex interaction network. The experimental and in silico results suggest two divergent evolutionary pathways: NMIIA and NMIIB evolved to possess S100A4-dependent and -independent regulations, respectively.

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“…These residues could be important to promote metastasis [32,33], and could also play some role in mediating interaction with certain partner proteins, like TG2, by fine tuning Ca 2+ -activation of S100A4, .as shown by our previous small angle X-ray scattering studies of the wild-type and its C-terminal deletion mutant protein [34]. Cysteine residues of S100A4 (Cys81 and Cys86), located in the hydrophobic binding pocket of S100A4 are crucial in binding to its well-described interaction partner NMIIA [16,26]. Mutating these cysteines to serines resulted in a 6.5 times higher K d value compared to the wild-type protein, indicating that presumably the binding region of S100A4 to TG2 could overlap with the NMIIA-binding hydrophobic binding pocket and the "waist" connecting the two pockets, though as noted above, the disordered Cterminal region could also have a minor role in the binding.…”

Section: Discussionmentioning

confidence: 92%

“…However, the presence of the above reactive lysine residue is not essential for the binding. Binding of S100A4 to NMIIA requires the presence of cysteine residues (Cys81 and Cys86) at the interaction surface [16,26]. Therefore the interaction of TG2 was investigated with a variant of S100A4 where these cysteine residues were mutated to serines (S100A4-Ser).…”

Section: S100a4 Is a Ca 2+ -Dependent Binding Partner Of Tg2mentioning

confidence: 99%

Metastasis-associated S100A4 is a specific amine donor and an activity-independent binding partner of transglutaminase-2

Biri

Kiss

Király

et al. 2015

Biochemical Journal

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Transglutaminase-2 (TG2) is best known as a Ca 2+ -dependent cross-linking enzyme; however, some of its extracellular matrix-related functions are independent from its catalytic activity and include matrix remodeling, adhesion and migration. S100A4 belongs to the Ca 2+ -binding EF-hand S100 protein family and acts both intra-and extracellularly through binding to various partners. It regulates cell migration and its overexpression is strongly associated with metastasis and poor survival in various cancers. TG2 has recently been suggested to mediate S100A4-dependent tumor cell migration. Here we provide evidence that S100A4 is an interacting partner and also specific amine donor of TG2. TG2 incorporates a glutamine donor peptide to Lys100 in the C-terminal random coil region of S100A4. Importantly, the enzyme activity is not necessary for the interaction: S100A4 also binds to TG2 in the presence of a specific inhibitor that keeps the enzyme in an open conformation, or to an enzymatically inactive mutant. We also found that S100A4 considerably enhances TG2-mediated adhesion of A431 epithelial carcinoma cells to the extracellular matrix. This role is independent of enzyme activity and requires the open conformation of TG2. We propose that S100A4 stabilizes the open conformation of TG2, which binds to its cell surface receptor in this state and increases cell adhesion.Summary: S100A4 and transglutaminase-2 have a role in metastasis. S100A4 is an interaction partner and specific amine substrate of transglutaminase-2, promoting its open conformation and leading to enhanced cell adhesion. Studying their interaction could contribute to the better understanding of metastasis.Running title: S100A4: substrate and binding partner of transglutaminase-2

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Asymmetric Mode of Ca2+-S100A4 Interaction with Nonmuscle Myosin IIA Generates Nanomolar Affinity Required for Filament Remodeling

Cited by 53 publications

References 42 publications

Crystal structure of the S100A4–nonmuscle myosin IIA tail fragment complex reveals an asymmetric target binding mechanism

Crystal structure of the S100A4–nonmuscle myosin IIA tail fragment complex reveals an asymmetric target binding mechanism

Structural determinants governing S100A4‐induced isoform‐selective disassembly of nonmuscle myosin II filaments

Metastasis-associated S100A4 is a specific amine donor and an activity-independent binding partner of transglutaminase-2

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