The 10.8-Å structure of Saccharomyces cerevisiae phosphofructokinase determined by cryoelectron microscopy: localization of the putative fructose 6-phosphate binding sites

Ruíz, Teresa; Mechin, I.; Bär, Jörg; Rypniewski, W.; Kopperschläger, Gèrhard; Radermacher, Michael

doi:10.1016/s1047-8477(03)00140-0

Cited by 32 publications

(48 citation statements)

References 42 publications

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“…Initially, the alignment was carried over an angular range of 180°in 5°a ngular increments. The averaged x, y, and z shifts were calculated from the 2D shifts of the three projections (2,28,30). The Euler angles for the volume rotation were determined from the angles obtained from the alignment of the (90°, 90°) projection.…”

Section: Methodsmentioning

confidence: 99%

Investigation of the Three-Dimensional Architecture of the Collagen Adhesin EmaA ofAggregatibacter actinomycetemcomitansby Electron Tomography

Yu¹,

Mintz²,

Ruíz

2009

J Bacteriol

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The periodontal pathogen Aggregatibacter actinomycetemcomitans displays on the bacterial surface a nonfimbrial adhesin, EmaA, which is required for collagen binding. In this study, electron tomography was used to characterize the three-dimensional (3D) architecture of this adhesin. The antenna-like surface appendages, corresponding to EmaA, were found to be composed of an ellipsoidal domain capping a rod-like domain that adopts either a straight or a bent conformation at various positions along the length. The most common flexible point along the length of the EmaA appendage was localized 29.4 nm away from the distal end. One-fifth of the appendages were straight and the remaining showed angles distributed between 140°and 170°at this location. Deletion analysis mapped this bend to amino acids 611 to 640 of the protein sequence. The 3D structure of the collagen binding domain of EmaA was generated by alignment and averaging of 9 subvolumes of the adhesin extracted from tomograms. The structure contains three subdomains: a globular structure with a diameter of ϳ5 nm and a cylindrical domain (ϳ4.4 nm by 5.8 nm) separated by a linker region with a diameter of ϳ3 nm, followed by a cylindrical domain (ϳ4.6 nm by 6.6 nm). This is the first 3D structure of a trimeric autotransporter protein of A. actinomycetemcomitans.

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Section: Methodsmentioning

confidence: 99%

Investigation of the Three-Dimensional Architecture of the Collagen Adhesin EmaA ofAggregatibacter actinomycetemcomitansby Electron Tomography

Yu¹,

Mintz²,

Ruíz

2009

J Bacteriol

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“…and a 14-m 2,048-by 2,048-pixel charge-coupled-device (CCD) camera (TVIPS, Gauting, Germany). The microscope was run under conditions identical to those that have been used in the past to obtain images that show Thon rings beyond 0.9-nm resolution in vitreous ice preparations (30). Images were recorded at an accelerating voltage of 100 kV and nominal magnifications in the range of ϫ40,000 to ϫ70,000 under low-dose conditions on either film (S0-163; Kodak) or the CCD camera.…”

Section: Methodsmentioning

confidence: 99%

Novel Surface Structures Are Associated with the Adhesion of Actinobacillus actinomycetemcomitans to Collagen

Ruíz¹,

Lenox

Radermacher³

et al. 2006

Infect Immun

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130

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Actinobacillus actinomycetemcomitans is a gram-negative, facultative, anaerobic bacterium that colonizes the human oral cavity and the upper respiratory tract. This bacterium is strongly associated with localized aggressive periodontitis and adult periodontitis and is the causative agent for other serious systemic infections. Recently, we have identified a protein, EmaA (extracellular matrix protein adhesin A), that mediates the adhesion of A. actinomycetemcomitans to collagen. The conserved sequence and predicted secondary structure suggest that EmaA is an orthologue of the Yersinia enterocolitica adhesin YadA. Electron microscopy examinations of A. actinomycetemcomitans have identified antenna-like protrusions associated with the surface of the bacterium. These structures are absent on emaA mutant strains and can be restored by transformation of the mutant strain with emaA in trans. The loss of these structures is associated with a decrease in the binding of this bacterium to collagen. The antenna-like structures are composed of a long rod that terminates in an ellipsoidal head region. The analysis of these structures using image processing techniques has provided an initial estimate of the overall dimensions, which suggests that the appendages are oligomeric structures formed by either three or four subunits. Together, the data suggest that emaA is required for the expression of novel appendages on the surface of A. actinomycetemcomitans that mediate the adhesion of the bacterium to collagen.

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“…The mammalian PFK was observed in various states of aggregation, while ScPFK proved suitable for single-particle EM reconstruction, revealing a relatively detailed (10.8 Å) structure of the octamer. [22][23][24] The crystal structure of the first eukaryotic PFK, from Trypanosoma brucei, displays features in common with both ATP-dependent and pyrophosphate-dependent bacterial PFKs but does not show gene duplication typical of eukaryotes. 25 A recent article describes the crystal structure of PFK from Pichia pastoris.…”

Section: Introductionmentioning

confidence: 99%

The Crystal Structures of Eukaryotic Phosphofructokinases from Baker's Yeast and Rabbit Skeletal Muscle

Banaszak

Mechin

Obmolova

et al. 2011

Journal of Molecular Biology

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The 10.8-Å structure of Saccharomyces cerevisiae phosphofructokinase determined by cryoelectron microscopy: localization of the putative fructose 6-phosphate binding sites

Cited by 32 publications

References 42 publications

Investigation of the Three-Dimensional Architecture of the Collagen Adhesin EmaA ofAggregatibacter actinomycetemcomitansby Electron Tomography

Investigation of the Three-Dimensional Architecture of the Collagen Adhesin EmaA ofAggregatibacter actinomycetemcomitansby Electron Tomography

Novel Surface Structures Are Associated with the Adhesion of Actinobacillus actinomycetemcomitans to Collagen

The Crystal Structures of Eukaryotic Phosphofructokinases from Baker's Yeast and Rabbit Skeletal Muscle

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