Solution structures of the C‐terminal headpiece subdomains of human villin and advillin, evaluation of headpiece F‐actin‐binding requirements

Vermeulen, Wim; Vanhaesebrouck, P.; Troys, Marleen Van; Verschueren, M Maykel; Fant, Franky; Goethals, Marc; Ampè, Christophe; Martins, José; Borremans, Frans

doi:10.1110/ps.03518104

Cited by 49 publications

(78 citation statements)

References 54 publications

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“…The core is further stabilized by van der Waals contacts between these Phes and the side chains of 10 conserved residues (Table 3) (32). Some of these contacts are also observed in the NMR structure 1VII, but a significant number (given in bold letters in Table 3) are not, again illustrating the differences between the x-ray and NMR structures.…”

Section: Resultsmentioning

confidence: 89%

“…Three highly conserved Phes, F6, F10, and F17 (31,32), form the bulk of the hydrophobic core. All are located within ␣1 and ␣2 and are presumably primarily responsible for the structural stability of this fragment (30).…”

Section: Resultsmentioning

confidence: 99%

“…1a). The survey of the villin and advillin family (32) shows that, with the exception of R14 and Q25, these nine amino acids can be replaced with equivalent ones as long as the contacting carbons are preserved. For example, the Leus at positions 20, 28, and 34 can also be a Thr, Ile, Met, Qln, Glu, or Phe (32).…”

Section: Resultsmentioning

confidence: 99%

“…As with R14, the amino nitrogen of Q25 is optimally oriented to donate a H bond to the peptide oxygens of A18 and L20. This residue is either a Gln, Arg, or Lys among the villin family, and the H-bond pattern is presumably preserved with either a Lys or Arg (32).…”

Section: Resultsmentioning

confidence: 99%

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High-resolution x-ray crystal structures of the villin headpiece subdomain, an ultrafast folding protein

Chiu

Kubelka

Herbst‐Irmer

et al. 2005

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

218

290

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The 35-residue subdomain of the villin headpiece (HP35) is a small ultrafast folding protein that is being intensely studied by experiments, theory, and simulations. We have solved the x-ray structures of HP35 and its fastest folding mutant [K24 norleucine (nL)] to atomic resolution and compared their experimentally measured folding kinetics by using laser temperature jump. The structures, which are in different space groups, are almost identical to each other but differ significantly from previously solved NMR structures. Hence, the differences between the x-ray and NMR structures are probably not caused by lattice contacts or crystal͞solution differences, but reflect the higher accuracy of the x-ray structures. The x-ray structures reveal important details of packing of the hydrophobic core and some additional features, such as crosshelical H bonds. Comparison of the x-ray structures indicates that the nL substitution produces only local perturbations. Consequently, the finding that the small stabilization by the mutation is completely reflected in an increased folding rate suggests that this region of the protein is as structured in the transition state as in the folded structure. It is therefore a target for engineering to increase the folding rate of the subdomain from Ϸ0.5 s ؊1 for the nL mutant to the estimated theoretical speed limit of Ϸ3 s ؊1 .kinetics ͉ temperature jump ͉ NMR R ecent developments in both experimental and computational methods have opened exciting new possibilities for combining experiments with all-atom molecular dynamics (MD) simulations of protein folding. The dramatic improvement in time resolution in protein folding kinetic studies brought about by the introduction of nanosecond optical triggering methods (1-4), together with the use of distributed computing, now make it possible to directly compare MD simulations with experiments on ultrafast folding proteins (5). Such simulations are important because, if accurate, they contain a complete atomic description of the folding mechanism. The computational cost of folding simulations demands small size and ultrafast folding. HP35 is a subdomain of the headpiece of actinbinding protein villin and is the smallest naturally occurring polypeptide that folds autonomously without any cofactor or disulfide bond (6). Unlike other ''miniproteins'' (7), HP35 is highly thermostable (6) and, as revealed by the NMR structure, globular with a well packed hydrophobic core (8). These properties resemble much larger single-domain helical proteins and have made HP36 (HP35 plus an N-terminal Met) the focus of several all-atom folding simulations (9-17), which have thus far been based on the NMR structure.Because of the delicate balance of stabilization forces involved in protein folding an accurate structure for the folded state is essential for MD simulations of folding and protein redesign calculations. We have previously shown that HP35 is one of the fastest folding proteins, with a folding rate (Ϸ0.2 s Ϫ1 ) approaching its theoretical limit (3 s Ϫ1 ...

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Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

High-resolution x-ray crystal structures of the villin headpiece subdomain, an ultrafast folding protein

Chiu

Kubelka

Herbst‐Irmer

et al. 2005

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

218

290

View full text Add to dashboard Cite

show abstract

“…This domain is highly conserved among the three different ABLIM isoforms (supplemental Fig. 1), as well as in ABLIM proteins from other species, including C. elegans (UNC-115) and Drosophila (D-UN-115), and has been shown to mediate actin binding (34,35). We thus directly tested the actin-binding properties of fulllength ABLIM-2 and -3, respectively, by conducting an actin co-sedimentation assay using in vitro-translated protein.…”

Section: Protein Expression and Subcellular Localization Of Ablim-2-mentioning

confidence: 99%

Two Novel Members of the ABLIM Protein Family, ABLIM-2 and -3, Associate with STARS and Directly Bind F-actin

Barrientos¹,

Frank

Kuwahara³

et al. 2007

Journal of Biological Chemistry

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In addition to regulating cell motility, contractility, and cytokinesis, the actin cytoskeleton plays a critical role in the regulation of transcription and gene expression. We have previously identified a novel muscle-specific actin-binding protein, STARS (striated muscle activator of Rho signaling), which directly binds actin and stimulates serum-response factor (SRF)-dependent transcription. To further dissect the STARS/SRF pathway, we performed a yeast two-hybrid screen of a skeletal muscle cDNA library using STARS as bait, and we identified two novel members of the ABLIM protein family, ABLIM-2 and -3, as STARSinteracting proteins. ABLIM-1, which is expressed in retina, brain, and muscle tissue, has been postulated to function as a tumor suppressor. ABLIM-2 and -3 display distinct tissue-specific expression patterns with the highest expression levels in muscle and neuronal tissue. Moreover, these novel ABLIM proteins strongly bind F-actin, are localized to actin stress fibers, and synergistically enhance STARS-dependent activation of SRF. Conversely, knockdown of endogenous ABLIM expression utilizing small interfering RNA significantly blunted SRF-dependent transcription in C2C12 skeletal muscle cells. These findings suggest that the members of the novel ABLIM protein family may serve as a scaffold for signaling modules of the actin cytoskeleton and thereby modulate transcription.

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The expanding superfamily of gelsolin homology domain proteins

et al. 2013

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The gelsolin homology (GH) domain has been found to date exclusively in actin-binding proteins. In humans, three copies of the domain are present in CapG, five copies in supervillin, and six copies each in adseverin, gelsolin, flightless I and the villins: villin, advillin and villin-like protein. Caenorhabditis elegans contains a four-GH-domain protein, GSNL-1. These architectures are predicted to have arisen from gene triplication followed by gene duplication to result in the six-domain protein. The subsequent loss of one, two or three domains produced the five-, four-, and three-domain proteins, respectively. Here we conducted BLAST and hidden Markov based searches of UniProt and NCBI databases to identify novel gelsolin domain containing proteins. The variety in architectures suggests that the GH domain has been tested in many molecular constructions during evolution. Of particular note is flightless-like I protein (FLIIL1) from Entamoeba histolytica, which combines a leucine rich repeats (LRR) domain, seven GH domains, and a headpiece domain, thus combining many of the features of flightless I with those of villin or supervillin. As such, the GH domain superfamily appears to have developed along complex routes. The distribution of these proteins was analyzed in the 343 completely sequenced genomes, mapped onto the tree of life, and phylogenetic trees of the proteins were constructed to gain insight into their evolution. © 2013 Wiley Periodicals, Inc.

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Solution structures of the C‐terminal headpiece subdomains of human villin and advillin, evaluation of headpiece F‐actin‐binding requirements

Cited by 49 publications

References 54 publications

High-resolution x-ray crystal structures of the villin headpiece subdomain, an ultrafast folding protein

High-resolution x-ray crystal structures of the villin headpiece subdomain, an ultrafast folding protein

Two Novel Members of the ABLIM Protein Family, ABLIM-2 and -3, Associate with STARS and Directly Bind F-actin

The expanding superfamily of gelsolin homology domain proteins

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