Structural characterization and Kemp eliminase activity of the Mycobacterium smegmatis Ketosteroid Isomerase

Thiolases catalyze the degradation and synthesis of 3-ketoacyl-CoA molecules. Here, the crystal structures of a T1-like thiolase (MSM-13 thiolase) fromMycobacterium smegmatisin apo and liganded forms are described. Systematic comparisons of six crystallographically independent unliganded MSM-13 thiolase tetramers (dimers of tight dimers) from three different crystal forms revealed that the two tight dimers are connected to a rigid tetramerization domainviaflexible hinge regions, generating an asymmetric tetramer. In the liganded structure, CoA is bound to those subunits that are rotated towards the tip of the tetramerization loop of the opposing dimer, suggesting that this loop is important for substrate binding. The hinge regions responsible for this rotation occur near Val123 and Arg149. The Lα1–covering loop–Lα2 region, together with the Nβ2–Nα2 loop of the adjacent subunit, defines a specificity pocket that is larger and more polar than those of other tetrameric thiolases, suggesting that MSM-13 thiolase has a distinct substrate specificity. Consistent with this finding, only residual activity was detected with acetoacetyl-CoA as the substrate in the degradative direction. No activity was observed with acetyl-CoA in the synthetic direction. Structural comparisons with other well characterized thiolases suggest that MSM-13 thiolase is probably a degradative thiolase that is specific for 3-ketoacyl-CoA molecules with polar, bulky acyl chains.

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Structural characterization of lipoarabinomannans from Mycobacterium tuberculosis and Mycobacterium smegmatis by ESI mass spectrometry

Journal of the American Society for Mass Spectrometry ◽

10.1016/j.jasms.2005.02.023 ◽

2005 ◽

Vol 16 (7) ◽

pp. 1109-1116 ◽

Cited By ~ 11

Author(s):

Christopher J. Petzold ◽

Leslie H. Stanton ◽

Julie A. Leary

Keyword(s):

Mass Spectrometry ◽

Mycobacterium Tuberculosis ◽

Structural Characterization ◽

Mycobacterium Smegmatis ◽

Esi Mass Spectrometry

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Phase behavior of cationic and anionic surfactants at low concentrations

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123878 ◽

1992 ◽

Vol 50 (1) ◽

pp. 694-695

Author(s):

E. Naranjo

Keyword(s):

Phase Behavior ◽

Structural Characterization ◽

Dynamic Systems ◽

Anionic Surfactants ◽

Intermolecular Forces ◽

Stable Form ◽

Surfactant Mixtures ◽

Low Concentrations ◽

Cationic And Anionic Surfactants ◽

General Method

Equilibrium vesicles, those which are the stable form of aggregation and form spontaneously on mixing surfactant with water, have never been demonstrated in single component bilayers and only rarely in lipid or surfactant mixtures. Designing a simple and general method for producing spontaneous and stable vesicles depends on a better understanding of the thermodynamics of aggregation, the interplay of intermolecular forces in surfactants, and an efficient way of doing structural characterization in dynamic systems.

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New ultrastructural findings about Borrelia burgdorferi by cryofixation, negative staining, thin sectioning and scanning techniques

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100160467 ◽

1990 ◽

Vol 48 (3) ◽

pp. 582-583

Author(s):

S. F. Hayes ◽

M. D. Corwin ◽

T. G. Schwan ◽

D. W. Dorward ◽

W. Burgdorfer

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Structural Characterization ◽

Negative Staining ◽

Low Speed ◽

Thin Sectioning ◽

Ultrastructural Findings

Characterization of Borrelia burgdorferi strains by means of negative staining EM has become an integral part of many studies related to the biology of the Lyme disease organism. However, relying solely upon negative staining to compare new isolates with prototype B31 or other borreliae is often unsatisfactory. To obtain more satisfactory results, we have relied upon a correlative approach encompassing a variety EM techniques, i.e., scanning for topographical features and cryotomy, negative staining and thin sectioning to provide a more complete structural characterization of B. burgdorferi.For characterization, isolates of B. burgdorferi were cultured in BSK II media from which they were removed by low speed centrifugation. The sedimented borrelia were carefully resuspended in stabilizing buffer so as to preserve their features for scanning and negative staining. Alternatively, others were prepared for conventional thin sectioning and for cryotomy using modified procedures. For thin sectioning, the fixative described by Ito, et al.

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