Structure and dynamics of the human muscle LIM protein

Schallus, Thomas; Fehér, Krisztina; Ulrich, Anne S.; Stier, Günter; Muhle‐Goll, Claudia

doi:10.1016/j.febslet.2009.02.021

Cited by 12 publications

(13 citation statements)

References 35 publications

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“…S9) and may cause aberrant induction of heart failure signalling. As all three HCM mutations are affecting conserved residues in the same region of the protein, namely the second zinc finger of the first LIM domain, it is likely that all three affect the protein structure [42] in a similar way and lead to partial protein unfolding as demonstrated previously for MLP C58G [4, 43]. These unfolded proteins will be recognised by protein quality control systems (as evidenced by the interaction of MLP C58G with Bag3 and the induction of Bag3 and associated heat shock proteins in our mouse model) and subsequently be targeted for degradation by the UPS.…”

Section: Discussionmentioning

confidence: 99%

“…These unfolded proteins will be recognised by protein quality control systems (as evidenced by the interaction of MLP C58G with Bag3 and the induction of Bag3 and associated heat shock proteins in our mouse model) and subsequently be targeted for degradation by the UPS. The mode of action appears to differ for DCM-associated mutations: K69R and G72R are located in the intrinsically unstructured glycine-rich region [42], hence an unfolded protein response in the presence of the mutations is unlikely. Instead, these mutations affect MLP's ability to inhibit PKC activity [6].…”

Section: Discussionmentioning

confidence: 99%

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Mutant Muscle LIM Protein C58G causes cardiomyopathy through protein depletion

Ehsan

Kelly

Hooper

et al. 2018

Journal of Molecular and Cellular Cardiology

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Cysteine and glycine rich protein 3 (CSRP3) encodes Muscle LIM Protein (MLP), a well-established disease gene for Hypertrophic Cardiomyopathy (HCM). MLP, in contrast to the proteins encoded by the other recognised HCM disease genes, is non-sarcomeric, and has important signalling functions in cardiomyocytes. To gain insight into the disease mechanisms involved, we generated a knock-in mouse (KI) model, carrying the well documented HCM-causing CSRP3 mutation C58G.In vivo phenotyping of homozygous KI/KI mice revealed a robust cardiomyopathy phenotype with diastolic and systolic left ventricular dysfunction, which was supported by increased heart weight measurements. Transcriptome analysis by RNA-seq identified activation of pro-fibrotic signalling, induction of the fetal gene programme and activation of markers of hypertrophic signalling in these hearts. Further ex vivo analyses validated the activation of these pathways at transcript and protein level. Intriguingly, the abundance of MLP decreased in KI/KI mice by 80% and in KI/+ mice by 50%. Protein depletion was also observed in cellular studies for two further HCM-causing CSRP3 mutations (L44P and S54R/E55G). We show that MLP depletion is caused by proteasome action. Moreover, MLP C58G interacts with Bag3 and results in a proteotoxic response in the homozygous knock-in mice, as shown by induction of Bag3 and associated heat shock proteins.In conclusion, the newly generated mouse model provides insights into the underlying disease mechanisms of cardiomyopathy caused by mutations in the non-sarcomeric protein MLP. Furthermore, our cellular experiments suggest that protein depletion and proteasomal overload also play a role in other HCM-causing CSPR3 mutations that we investigated, indicating that reduced levels of functional MLP may be a common mechanism for HCM-causing CSPR3 mutations.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Mutant Muscle LIM Protein C58G causes cardiomyopathy through protein depletion

Ehsan

Kelly

Hooper

et al. 2018

Journal of Molecular and Cellular Cardiology

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“…To demonstrate the difference in disorder predictions between servers, we submitted cardiac Muscle LIM Protein (MLP) ( Figure 2 ) to various servers ( Table 2 ). This protein is known to contain a long disordered region within the central region, similar to other members of the CRP family [ 53 , 54 ]. As with most, if not all, proteins, both the N- and C-termini contain some degree of disorder.…”

Section: Disorder Prediction Methodologies and Publicly Available mentioning

confidence: 99%

Disorder Prediction Methods, Their Applicability to Different Protein Targets and Their Usefulness for Guiding Experimental Studies

Atkins

Boateng

Sørensen

et al. 2015

IJMS

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The role and function of a given protein is dependent on its structure. In recent years, however, numerous studies have highlighted the importance of unstructured, or disordered regions in governing a protein’s function. Disordered proteins have been found to play important roles in pivotal cellular functions, such as DNA binding and signalling cascades. Studying proteins with extended disordered regions is often problematic as they can be challenging to express, purify and crystallise. This means that interpretable experimental data on protein disorder is hard to generate. As a result, predictive computational tools have been developed with the aim of predicting the level and location of disorder within a protein. Currently, over 60 prediction servers exist, utilizing different methods for classifying disorder and different training sets. Here we review several good performing, publicly available prediction methods, comparing their application and discussing how disorder prediction servers can be used to aid the experimental solution of protein structure. The use of disorder prediction methods allows us to adopt a more targeted approach to experimental studies by accurately identifying the boundaries of ordered protein domains so that they may be investigated separately, thereby increasing the likelihood of their successful experimental solution.

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“…At least three regions of titin form complexes with other proteins that are implicated in signaling. In titin’s Z-line region, repeats Z1-Z2 interacts with T-cap/telethonin,4 which itself interacts with a potassium channel subunit,5 myostatin (a muscle growth factor),6 and the muscle LIM protein (MLP) 7. Z-line repeat Z4, and the 700 kDa alternative titin isoform “novex-3 titin” (located in the I-band near the Z-line) interact with obscurin, a ~700 kDa protein that is involved in regulating Rho-like GTPases.…”

Section: Introductionmentioning

confidence: 99%

Extensive and Modular Intrinsically Disordered Segments in C. elegans TTN-1 and Implications in Filament Binding, Elasticity and Oblique Striation

Forbes

Flaherty

et al. 2010

Journal of Molecular Biology

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TTN-1, a predicted titin-like protein in C. elegans, is encoded by a single gene, and consists of multiple Ig and Fn3 domains, a protein kinase domain and several regions containing tandem short repeat sequences. We have characterized TTN-1’s sarcomere distribution, protein interaction with key myofibrillar proteins as well as the conformation malleability of representative motifs of five classes of short repeats. We report that two antibodies developed to portions of TTN-1 detect a ~2 MDa polypeptide on western blots. In addition, by immunofluorescence staining, both of these antibodies localize to the I-band and may extend into the outer edge of the A-band in the obliquely striated muscle of the nematode. Six different 300 residue segments of TTN-1 were shown to variously interact with actin and/or myosin in vitro. Conformations of synthetic peptides of representative copies of each of the five classes of repeats: 39-mer PEVT, 51-mer CEEEI, 42-mer AAPLE, 32-mer BLUE and 30-mer DispRep were investigated by circular dichroism at different temperatures, ionic strengths and solvent polarities. The PEVT, CEEEI, DispRep and AAPLE peptides displayed a combination of a polyproline II helix and an unordered structure in aqueous solution and converts in trifluoroethanol to α-helix (PEVT, DispRep) and β-turn (AAPLE) structures, respectively. The octads in BLUE motifs form unstable α-helix-unstructured coil in aqueous solution and negligible heptad-based coiled-coils, as predicted based on sequence analysis. The α-helical structure, as modeled by threading and molecular dynamics simulations, tends to form helical bundles and crosses based on its 8-4-2-2 hydrophobic helical patterns and charge arrays on its surface. Our finding indicates that APPLE, PEVT, DispRep regions are all intrinsically disordered and highly reminiscent of the conformational malleability and elasticity of vertebrate titin PEVK segments. The presence of long, modular and unstable α-helical oligomerization domains in the BLUE region of TTN-1 could oligomerize and bundle TTN-1 and stabilize oblique striation of the sarcomere.

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Structure and dynamics of the human muscle LIM protein

Cited by 12 publications

References 35 publications

Mutant Muscle LIM Protein C58G causes cardiomyopathy through protein depletion

Mutant Muscle LIM Protein C58G causes cardiomyopathy through protein depletion

Disorder Prediction Methods, Their Applicability to Different Protein Targets and Their Usefulness for Guiding Experimental Studies

Extensive and Modular Intrinsically Disordered Segments in C. elegans TTN-1 and Implications in Filament Binding, Elasticity and Oblique Striation

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