Atomic model of the actin filament

Holmes, Kenneth C.; Popp, David; Gebhard, Werner; Kabsch, Wolfgang

doi:10.1038/347044a0

Cited by 1,524 publications

(1,343 citation statements)

References 43 publications

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“…Evidence is presented that photoactivation of the azidobenzoyl moiety results in formation of a didehydroazepine-mediated reaction with an €-amino group, yielding an intermolecular cross-link that is slowly hydrolyzed in weak acid. This cross-link, as well as one described previously, can be accommodated within, and therefore supports, the F-actin model of Holmes et al (1990). Determination of the atomic coordinates of the relevant amino acid side chains will permit refinement of the spatial localization of the cross-links.…”

Section: Actin-actin Contact Pointssupporting

confidence: 70%

“…In the model of Holmes et al (1990), the a-carbons of these two side chains are about 19 A apart in adjacent monomers along the genetic heIix; thus, this cross-link (phenylene-dimaleimide) must extend across the central axis of the polymer. In other words, if one considers F-actin to be composed of two strands, this cross-link connects Cys-374 of a monomer in one strand with Lys-191 of a monomer in the other strand.…”

Section: Structural Implicationsmentioning

confidence: 99%

“…This loop is a major contributor to the DNase I binding site of G-actin and is an important point of contact between actin monomers in the Holmes et al (1990) model of F-actin. Lys-113 is part of an a-helix on the surface of a domain that contains the amino-terminus, the carboxyl-terminus, and residues -75 to -135.…”

Section: The Gln-ll:lys-113 Cross-linkmentioning

confidence: 99%

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Gln‐41 is intermolecularly cross‐linked to Lys‐113 in F‐actin by N‐(4‐azidobenzoyl)‐putrescine

Hegyi¹,

Michel²,

Shabanowitz³

et al. 1992

Protein Science

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The bifunctional reagent N-(4-azidobenzoyl)-putrescine was synthesized and covalently bound to rabbit skeletal muscle actin. The incorporation was mediated by guinea pig liver transglutaminase under conditions similar to those described by Takashi (1988, Biochemistry 27, 938-943); up to 0.5 M/M were incorporated into G-actin, whereas F-actin was refractory to incorporation. Peptide fractionation showed that at least 90% of the label was bound to Gln-41. The labeled G-actin was polymerized, and irradiation of the F-actin led to covalent intermolecular cross-linking. A cross-linked peptide complex was isolated from a tryptic digest of the cross-linked actin in which digestion was limited to arginine; sequence analysis as well as mass spectrometry indicated that the linked peptides contained residues 40-62 and residues 96-116, and that the actual cross-link was between Gln-41 and Lys-113. Thus the y-carboxyl group of Gln-41 must be within 10.7 A of the side chain (probably the amino group) of Lys-I13 in an adjacent actin monomer. In the atomic model for F-actin proposed by Holmes et al. (1990, Nuture 347, 44-49), the a-carbons of these residues in adjacent monomers along the two-start helices are sufficiently close to permit cross-linking of their side chains, and, pending atomic resolution of the side chains, the results presented here seem to support the proposed model.

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Section: Actin-actin Contact Pointssupporting

confidence: 70%

Section: Structural Implicationsmentioning

confidence: 99%

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Gln‐41 is intermolecularly cross‐linked to Lys‐113 in F‐actin by N‐(4‐azidobenzoyl)‐putrescine

Hegyi¹,

Michel²,

Shabanowitz³

et al. 1992

Protein Science

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“…3), similar as F-actin (7-9 nm) [45,46]. These results above indicate that the prokaryotic expressed PEAc1-GFP fusion proteins were able to polymerize into F-actin-like structures in vitro.…”

Section: Peac1-gfp Fusion Proteins Can Polymerize Into a Filamentousmentioning

confidence: 79%

Soluble expression and characterization of a GFP-fused pea actin isoform (PEAc1)

Liu

Zhang

et al. 2004

Cell Res

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A pea actin isoform PEAc1 with green fluorescent protein (GFP) fusion to its C-terminus and His-tag to its Nterminus, was expressed in prokaryotic cells in soluble form, and highly purified with Ni-Chelating Sepharose TM Fast Flow column. The purified fusion protein (PEAc1-GFP) efficiently inhibited DNase I activities before polymerization, and activated the myosin Mg-ATPase activities after polymerization. The PEAc1-GFP also polymerized into green fluorescent filamentous structures with a critical concentration of 0.75 µM. These filamentous structures were labeled by TRITC-phalloidin, a specific agent for staining actin microfilaments, and identified as having 9 nm diameters by negative staining. These results indicated that PEAc1 preserved the essential characteristics of actin even with His-tag and GFP fusion, suggesting a promising potential to use GFP fusion protein in obtainning soluble plant actin isoform to analyze its physical and biochemical properties in vitro. The PEAc1-GFP was also expressed in tobacco BY2 cells, which offers a new pathway for further studying its distribution and function in vivo.

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“…Atomic resolution of the actin structure [1,2] and the threeciimensional atomic model of F-actin decorated with rabbit ~aymotryptic-S1 [3] or Dictyostelium myosin S1 [4] revealed I ~cations of the myosin head on F-actin. This molecular model f the actin-myosin complex derived from the X-ray structure l zveals a close contact between a myosin subfragment-I and 1 ~o actin monomers.…”

Section: ~ Introductionmentioning

confidence: 99%

Two identical hydrophobic clusters are present on the same actin monomer: interaction between one myosin subfragment‐1 and two actin monomers

Labbé¹,

Boyer²,

Benyamin³

1995

FEBS Letters

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Two-dimensional hydrophobic clusters analysis (IICA) was used to compare the distribution of hydrophobic clusters along various actin sequence. HCA-deduced patterns were not altered by amino-acid variations throughout the evolution of actin and we observed similar hydrophobic motifs comprising myosin subfragment-I ATP-independent binding sites. HCA suggested the presence of two groups of identical hydrophobic motifs (A~ and A2) which bound on each side of the S1 (63 kDa-31 kDa) connecting segment in relation with two actin monomers. This connection is important in communications between actin-and nucleotide-binding sites. We postulate that some relation and message between the two motifs A~ and A 2 take place through myosin subfragment-I (63 kDa-31 kDa) connecting segment.1~ ey words'," Acto-myosin; Hydrophobic cluster; Structure comparison 1~ IntroductionIdentification of the actin-myosin interface is essential in t nderstanding the cyclical enzymatic and mechanical process c f myofibrillar contraction.Atomic resolution of the actin structure [1,2] and the threeciimensional atomic model of F-actin decorated with rabbit ~aymotryptic-S1 [3] or Dictyostelium myosin S1 [4] revealed I ~cations of the myosin head on F-actin. This molecular model f the actin-myosin complex derived from the X-ray structure l zveals a close contact between a myosin subfragment-I and 1 ~o actin monomers. A first contact includes carboxyl residues ,2, 3, 4, 24, 25, 99 and 100. In a second contact, exposed actin 1 ydrophobic residues 144, 341,345,349 and 352 are concerned. "he third contact involves proline residues (332-333) of actin. ~11 of these contacts involve a single monomer. A second mon-~,mer is close to the myosin heavy chain and the interaction i ~cludes the cz-helix formed by actin residues Trp79 to Asn92 flat formed a weak binding contact and was cross-linked [5,6].The Taylor-Amos model [7] of acto-S1 suggests that S1 has n extensive contact with an actin monomer (acl) and a weaker ,. ontact with an adjacent actin monomer (ac2); and two differ-,, nt rigor complexes were suggested [8,9], with SI binding to :-actin occurring in two steps with actin monomers. Using .,uccessively EDC and DMS, we previously cross-linked two lnonomers to 20-kDa and 50-kDa skeletal S1 fragments, re-: Corresponding author. Fax: (33) (67) 60 94 78.spectively [10], and in vitro studies showed that subfragment-I alone can produce the sliding movement of the actin filament [111.Chemical cross-linking [12], NMR [13,14] and immunochemical studies [15,16,[17][18][19][20] have pinpointed actin segments in subdomain-1, residues 1 28, 96-103, 112-125, 338 348 and 360-372 which are able to bind with the myosin head.Furthermore, the (27 kDa-50 kDa-20 kDa) trypsin split myosin subfragment-1, which could no longer be activated by actin, did not bind to the two sites located in the 96-125 region, but it still interacted with the 338-348 and 360--372 segments [191. The presence of hydrophobic interactions in the actin myosin complex have already been sugge...

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Atomic model of the actin filament

Cited by 1,524 publications

References 43 publications

Gln‐41 is intermolecularly cross‐linked to Lys‐113 in F‐actin by N‐(4‐azidobenzoyl)‐putrescine

Gln‐41 is intermolecularly cross‐linked to Lys‐113 in F‐actin by N‐(4‐azidobenzoyl)‐putrescine

Soluble expression and characterization of a GFP-fused pea actin isoform (PEAc1)

Two identical hydrophobic clusters are present on the same actin monomer: interaction between one myosin subfragment‐1 and two actin monomers

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