SEPARATION OF MITOCHONDRIAL MEMBRANES OF <i>NEUROSPORA CRASSA  </i>

Cassady, W. E.; Wagner, Robert P.

doi:10.1083/jcb.49.2.536

Cited by 28 publications

(14 citation statements)

References 20 publications

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“…Separation of the submitochondrial fractions after digitonin treatment was accomplished with a discontinuous sucrose gradient as previously described (8), with the modification that 1 .0 ml of 1 .9 M sucrose was used as a cushion ; 1 .5 ml of 1 .0 M sucrose was layered above this, and a 2 .0 ml sample of digitonintreated mitochondria was layered on top . The gradient fractions observed, after centrifugation, are shown diagrammatically in Fig .…”

Section: Digitonin Treatment and Submitochondrial Fractionationmentioning

confidence: 99%

“…The B 1 and B2 fractions were fixed in 2 % buffered glutaraldehyde, pH 7 .2, postfixed in 1 % osmium tetroxide for I hr, stained in 0 .5% uranyl acetate, and prepared for electron microscopy with a Siemens Elmiskop 1 as previously described (8) .…”

Section: Electron Microscopymentioning

confidence: 99%

“…Mitochondria from wild type Neurospora crassa strain LSDT(1969A) were prepared by the sandground method previously described (8) . The crude mitochondrial fraction was washed once with 0 .25 M sucrose, 0 .15% bovine serum albumin (BSA), then centrifuged at 37,000 g in a Sorvall SS-34 rotor .…”

Section: Purification Of Mitochondriamentioning

confidence: 99%

“…Malate dehydrogenase (MDH) was assayed by the method of Ochoa (13) . SCCR was assayed by the method of Tisdale (14), using a modification by Cassady and Wagner (8) . KH was assayed by the method of Ghosh and Forrest (15) .…”

Section: Electron Microscopymentioning

confidence: 99%

“…Submitochondrial localization of these enzymes has been dependent upon the development of enzymatic markers for outer membrane (1,6), matrix (3), and inner membrane (7) . Lately, in this laboratory, Cassady and Wagner (8) as an outer membrane marker, and succinatecytochrome c reductase (SCCR) as an inner membrane marker . In this study, digitonin subfractionation has been used to uncover evidence that a group of four enzymes catalyzing the over-all biosynthesis of two branched chain amino acids, isoleucine and valine, from pyruvate-14 C in isolated Neurospora mitochondria (9), behave as soluble enzymes loosely associated in the mitochondrial matrix.…”

Section: Introductionmentioning

confidence: 99%

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SEPARATION OF MITOCHONDRIAL MEMBRANES OF NEUROSPORA CRASSA

Cassady

Leiter

Bergquist

et al. 1972

The Journal of Cell Biology

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Separation of Neurospora mitochondrial outer membranes from the inner membrane/matrix fraction was effected by digitonin treatment and discontinuous density gradient centrifugation. The solubilization of four isoleucine-valine biosynthetic enzymes was studied as a function of digitonin concentration and time of incubation in the detergent. The kinetics of the appearance of valine biosynthetic function in fractions outside of the inner membrane/matrix fraction, coupled with enzyme solubilization patterns similar to that for the matrix marker, mitochondrial malate dehydrogenase, indicate that the four isoleucine-valine pathway enzymes are localized in the mitochondrial matrix.

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Section: Digitonin Treatment and Submitochondrial Fractionationmentioning

confidence: 99%

Section: Electron Microscopymentioning

confidence: 99%

Section: Purification Of Mitochondriamentioning

confidence: 99%

Section: Electron Microscopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

SEPARATION OF MITOCHONDRIAL MEMBRANES OF NEUROSPORA CRASSA

Cassady

Leiter

Bergquist

et al. 1972

The Journal of Cell Biology

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Subcellular localization of the first three enzymes of the ommochrome synthetic pathway in Drosophila melanogaster

1974

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Proteome analysis of mitochondrial outer membrane from Neurospora crassa

Schmitt

Prokisch

Schlunck³

et al. 2006

Proteomics

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The mitochondrial outer membrane mediates numerous interactions between the metabolic and genetic systems of mitochondria and the rest of the eukaryotic cell. We performed a proteomic study to discover novel functions of components of the mitochondrial outer membrane. Proteins of highly pure outer membrane vesicles (OMV) from Neurospora crassa were identified by a combination of LC-MS/MS of tryptic peptide digests and gel electrophoresis of solubilized OMV proteins, followed by their identification using MALDI-MS PMF. Among the 30 proteins found in at least three of four separate analyses were 23 proteins with known functions in the outer membrane. These included components of the import machinery (the TOM and TOB complexes), a pore-forming component (porin), and proteins that control fusion and fission of the organelle. In addition, proteins playing a role in various biosynthetic pathways, whose intracellular location had not been established previously, could be localized to the mitochondrial outer membrane. Thus, the proteome of the outer membrane can help in identifying new mitochondria-related functions.

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SEPARATION OF MITOCHONDRIAL MEMBRANES OF NEUROSPORA CRASSA

Cited by 28 publications

References 20 publications

SEPARATION OF MITOCHONDRIAL MEMBRANES OF NEUROSPORA CRASSA

SEPARATION OF MITOCHONDRIAL MEMBRANES OF NEUROSPORA CRASSA

Subcellular localization of the first three enzymes of the ommochrome synthetic pathway in Drosophila melanogaster

Proteome analysis of mitochondrial outer membrane from Neurospora crassa

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