TY - JOUR
T1 - Cryo-electron tomography of Neurospora mitochondria
AU - Nicastro, Daniela
AU - Frangakis, Achilleas S.
AU - Typke, Dieter
AU - Baumeister, Wolfgang
N1 - Funding Information:
We thank W. Neupert, S. Nussberger, P. Heckmeyer, and M. Braun for supplying us with conidia of Neurospora crassa and introducing us to their isolation protocol of mitochondria as well as for helpful discussions. We are also grateful to D. Brdiczka for helpful advice and to M. Kania for carefully reading the manuscript. This work was supported by a grant from the EU (Contract BIO4 CT960099).
PY - 2000/2
Y1 - 2000/2
N2 - Cryo-electron tomography was used to study the structural organization of whole frozen-hydrated mitochondria from Neurospora crassa. Unlike mitochondria from many other species and tissues, in this case the cristae form a three-dimensional network of interconnected lamellae. Basically, the three-dimensional structure of ice-embedded mitochondria from this species is consistent with previous descriptions of mitochondria prepared by chemical fixation and resin embedding. Nonetheless, ice-embedded mitochondria display some important differences: the outer surface of the mitochondria was found to be rather smooth, the intermembrane space was constant in width, and distinct contact sites between the membranes were clearly revealed. Furthermore ATP synthase particles on the outer surface of an 'inside-out vesicle' were visible in 3-D reconstructions. Thus, cryo-electron tomography can provide detailed insights into these organelles with minimal perturbations of the physiological state. This indicates that it is a realistic goal to achieve 'molecular resolution' with rather large biological specimens in the near future, ultimately allowing the identification and localization of macromolecules in their cellular context. (C) 2000 Academic Press.
AB - Cryo-electron tomography was used to study the structural organization of whole frozen-hydrated mitochondria from Neurospora crassa. Unlike mitochondria from many other species and tissues, in this case the cristae form a three-dimensional network of interconnected lamellae. Basically, the three-dimensional structure of ice-embedded mitochondria from this species is consistent with previous descriptions of mitochondria prepared by chemical fixation and resin embedding. Nonetheless, ice-embedded mitochondria display some important differences: the outer surface of the mitochondria was found to be rather smooth, the intermembrane space was constant in width, and distinct contact sites between the membranes were clearly revealed. Furthermore ATP synthase particles on the outer surface of an 'inside-out vesicle' were visible in 3-D reconstructions. Thus, cryo-electron tomography can provide detailed insights into these organelles with minimal perturbations of the physiological state. This indicates that it is a realistic goal to achieve 'molecular resolution' with rather large biological specimens in the near future, ultimately allowing the identification and localization of macromolecules in their cellular context. (C) 2000 Academic Press.
KW - ATP synthase
KW - Cellular tomography
KW - Contact sites
KW - Cryo-electron microscopy
KW - Frozen-hydrated specimens
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U2 - 10.1006/jsbi.1999.4204
DO - 10.1006/jsbi.1999.4204
M3 - Article
C2 - 10675296
AN - SCOPUS:0034010882
SN - 1047-8477
VL - 129
SP - 48
EP - 56
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 1
ER -