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Title: Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott)

Abstract

Polyclonal and monoclonal antibodies that recognize the 35-, 36- and 37-kDa alternative oxidase proteins of Sauromatum guttatum (Schott) were used to isolate a cDNA clone, pAOSG81, from an S. guttatum cDNA expression library. A fusion protein with an apparent molecular mass of 48 kDa was expressed from a pUC119 derivative of pAOSG81 (pAOSG81-119) in Escherichia coli cells and was recognized by the monoclonal antibodies. When the in vitro translated and immunoprecipitated products made form mRNA hybrid-selected by pAOSG81 were analyzed, a single band corresponding to a protein with an apparent molecular mass of 42kDa was observed. DNA sequence characterization showed that pAOSG81 contains the entire coding region of a protein with a calculated molecular mass of 38.9 kDa, a putative 63-amino acid transit peptide, and a 9-amino acid match to the authentic N-terminal sequence of the 36-kDa alternative oxidase protein. Analyses of the deduced amino acid sequence indicate: (1) that the transit peptide is predicted to form amphiphilic helices, and (2) that three regions of the processed protein are likely to form transmembrane {alpha}-helices. The authors conclude from these data that pALSG81 represents a nuclear gene, aox1, encoding a precursor protein of one or more of the alternative oxidase proteinsmore » of S. guttatum.« less

Authors:
;  [1]
  1. Michigan State Univ., East Lansing (United States)
Publication Date:
OSTI Identifier:
5026736
DOE Contract Number:  
AC02-76ER01338
Resource Type:
Journal Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America; (United States)
Additional Journal Information:
Journal Volume: 88:6; Journal ID: ISSN 0027-8424
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CYTOCHROME OXIDASE; AMINO ACID SEQUENCE; PLANT CELLS; BIOLOGICAL PATHWAYS; RECOMBINANT DNA; DNA SEQUENCING; DNA HYBRIDIZATION; ESCHERICHIA COLI; FLOWERS; GENE REGULATION; MESSENGER-RNA; MITOCHONDRIA; MOLECULAR WEIGHT; MONOCLONAL ANTIBODIES; ANTIBODIES; BACTERIA; CELL CONSTITUENTS; DNA; ENZYMES; HAEM DEHYDROGENASES; HYBRIDIZATION; MICROORGANISMS; MOLECULAR STRUCTURE; NUCLEIC ACIDS; ORGANIC COMPOUNDS; OXIDOREDUCTASES; RNA; STRUCTURAL CHEMICAL ANALYSIS; 550200* - Biochemistry

Citation Formats

Rhoads, D M, and McIntosh, L. Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott). United States: N. p., 1991. Web. doi:10.1073/pnas.88.6.2122.
Rhoads, D M, & McIntosh, L. Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott). United States. https://doi.org/10.1073/pnas.88.6.2122
Rhoads, D M, and McIntosh, L. 1991. "Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott)". United States. https://doi.org/10.1073/pnas.88.6.2122.
@article{osti_5026736,
title = {Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott)},
author = {Rhoads, D M and McIntosh, L},
abstractNote = {Polyclonal and monoclonal antibodies that recognize the 35-, 36- and 37-kDa alternative oxidase proteins of Sauromatum guttatum (Schott) were used to isolate a cDNA clone, pAOSG81, from an S. guttatum cDNA expression library. A fusion protein with an apparent molecular mass of 48 kDa was expressed from a pUC119 derivative of pAOSG81 (pAOSG81-119) in Escherichia coli cells and was recognized by the monoclonal antibodies. When the in vitro translated and immunoprecipitated products made form mRNA hybrid-selected by pAOSG81 were analyzed, a single band corresponding to a protein with an apparent molecular mass of 42kDa was observed. DNA sequence characterization showed that pAOSG81 contains the entire coding region of a protein with a calculated molecular mass of 38.9 kDa, a putative 63-amino acid transit peptide, and a 9-amino acid match to the authentic N-terminal sequence of the 36-kDa alternative oxidase protein. Analyses of the deduced amino acid sequence indicate: (1) that the transit peptide is predicted to form amphiphilic helices, and (2) that three regions of the processed protein are likely to form transmembrane {alpha}-helices. The authors conclude from these data that pALSG81 represents a nuclear gene, aox1, encoding a precursor protein of one or more of the alternative oxidase proteins of S. guttatum.},
doi = {10.1073/pnas.88.6.2122},
url = {https://www.osti.gov/biblio/5026736}, journal = {Proceedings of the National Academy of Sciences of the United States of America; (United States)},
issn = {0027-8424},
number = ,
volume = 88:6,
place = {United States},
year = {Fri Mar 15 00:00:00 EST 1991},
month = {Fri Mar 15 00:00:00 EST 1991}
}