Details
| Original language | English |
|---|---|
| Pages (from-to) | 878-885 |
| Number of pages | 8 |
| Journal | European Journal of Biochemistry |
| Volume | 228 |
| Issue number | 3 |
| Publication status | Published - Mar 1995 |
| Externally published | Yes |
Abstract
Analysis of cytochrome c reductase from potato by Tricine/SDS/PAGE reveals 10 bands representing 10 different subunits. In comparison to glycine/SDS/PAGE one additional small protein becomes visible, whereas the three large core proteins are not resolved. The identity of the subunits was determined by immunoblotting and direct sequence determination. Sequence data for the novel small component were used to derive oligonuleotides for probing a potato cDNA‐library and isolating corresponding clones. The newly identified subunit is a 6.7‐kDa protein, that exhibits significant sequence similatity to a 8.5‐kDa subunit of cytochrome c reductase from yeast and the 6.5‐kDa iron‐sulfur‐protein‐binding factor from the equivalent enzyme complex from beef. Also the potato 6.7‐kDa subunit can be dissociated from the cytochrome c reductase complex together with the iron‐sulfur protein. To address the question of whether three or two core subunits occur simultaneously in monomeric cytochrome c reductase complexes from potato, a peptide‐specific antibody was generated. The antiserum is capable of discriminating between the 55‐kDa and 53‐kDa core proteins, which can be separated by glycine/SDS/PAGE and which were previously found to be structurally related. Immunoprecipitations of isolated cytochrome c reductase from potato using this antibody revealed an enzyme complex containing only two core proteins. The simultaneous occurrence of only two core subunits was confirmed by a comparison of the molecular masses of cytochrome c reductase from potato and beef by blue‐native‐gel electrophoresis. Hence the cytochrome c reductase complexes from potato, beef and yeast have a very conserved subunit composition. The evolutionary implications of these findings are discussed.
Keywords
- Cytochrome c reductase, mitochondrial processing peptidase, mitochondrial protein import, respiration, Solanum tuberosum
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
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In: European Journal of Biochemistry, Vol. 228, No. 3, 03.1995, p. 878-885.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Cytochrome c Reductase from Potato Does not Comprise Three Core Proteins but Contains an Additional Low‐Molecular‐Mass Subunit
AU - Jänsch, Lothar
AU - Kruft, Volker
AU - Schmitz, Udo
AU - Braun, Hans‐Peter
PY - 1995/3
Y1 - 1995/3
N2 - Analysis of cytochrome c reductase from potato by Tricine/SDS/PAGE reveals 10 bands representing 10 different subunits. In comparison to glycine/SDS/PAGE one additional small protein becomes visible, whereas the three large core proteins are not resolved. The identity of the subunits was determined by immunoblotting and direct sequence determination. Sequence data for the novel small component were used to derive oligonuleotides for probing a potato cDNA‐library and isolating corresponding clones. The newly identified subunit is a 6.7‐kDa protein, that exhibits significant sequence similatity to a 8.5‐kDa subunit of cytochrome c reductase from yeast and the 6.5‐kDa iron‐sulfur‐protein‐binding factor from the equivalent enzyme complex from beef. Also the potato 6.7‐kDa subunit can be dissociated from the cytochrome c reductase complex together with the iron‐sulfur protein. To address the question of whether three or two core subunits occur simultaneously in monomeric cytochrome c reductase complexes from potato, a peptide‐specific antibody was generated. The antiserum is capable of discriminating between the 55‐kDa and 53‐kDa core proteins, which can be separated by glycine/SDS/PAGE and which were previously found to be structurally related. Immunoprecipitations of isolated cytochrome c reductase from potato using this antibody revealed an enzyme complex containing only two core proteins. The simultaneous occurrence of only two core subunits was confirmed by a comparison of the molecular masses of cytochrome c reductase from potato and beef by blue‐native‐gel electrophoresis. Hence the cytochrome c reductase complexes from potato, beef and yeast have a very conserved subunit composition. The evolutionary implications of these findings are discussed.
AB - Analysis of cytochrome c reductase from potato by Tricine/SDS/PAGE reveals 10 bands representing 10 different subunits. In comparison to glycine/SDS/PAGE one additional small protein becomes visible, whereas the three large core proteins are not resolved. The identity of the subunits was determined by immunoblotting and direct sequence determination. Sequence data for the novel small component were used to derive oligonuleotides for probing a potato cDNA‐library and isolating corresponding clones. The newly identified subunit is a 6.7‐kDa protein, that exhibits significant sequence similatity to a 8.5‐kDa subunit of cytochrome c reductase from yeast and the 6.5‐kDa iron‐sulfur‐protein‐binding factor from the equivalent enzyme complex from beef. Also the potato 6.7‐kDa subunit can be dissociated from the cytochrome c reductase complex together with the iron‐sulfur protein. To address the question of whether three or two core subunits occur simultaneously in monomeric cytochrome c reductase complexes from potato, a peptide‐specific antibody was generated. The antiserum is capable of discriminating between the 55‐kDa and 53‐kDa core proteins, which can be separated by glycine/SDS/PAGE and which were previously found to be structurally related. Immunoprecipitations of isolated cytochrome c reductase from potato using this antibody revealed an enzyme complex containing only two core proteins. The simultaneous occurrence of only two core subunits was confirmed by a comparison of the molecular masses of cytochrome c reductase from potato and beef by blue‐native‐gel electrophoresis. Hence the cytochrome c reductase complexes from potato, beef and yeast have a very conserved subunit composition. The evolutionary implications of these findings are discussed.
KW - Cytochrome c reductase
KW - mitochondrial processing peptidase
KW - mitochondrial protein import
KW - respiration
KW - Solanum tuberosum
UR - http://www.scopus.com/inward/record.url?scp=0028934057&partnerID=8YFLogxK
U2 - 10.1111/j.1432-1033.1995.tb20335.x
DO - 10.1111/j.1432-1033.1995.tb20335.x
M3 - Article
C2 - 7737189
AN - SCOPUS:0028934057
VL - 228
SP - 878
EP - 885
JO - European Journal of Biochemistry
JF - European Journal of Biochemistry
SN - 0014-2956
IS - 3
ER -