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

Journal Article · Fri Mar 15 04:00:00 EST 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.88.6.2122· OSTI ID:5026736

Rhoads, D M; McIntosh, L ^[1]

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1. Michigan State Univ., East Lansing (United States)

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.

• DOE Contract Number: AC02-76ER01338
• OSTI ID: 5026736
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 88:6; ISSN PNASA; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANTIBODIES
BACTERIA
BIOLOGICAL PATHWAYS
CELL CONSTITUENTS
CYTOCHROME OXIDASE
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
ENZYMES
ESCHERICHIA COLI
FLOWERS
GENE REGULATION
HAEM DEHYDROGENASES
HYBRIDIZATION
MESSENGER-RNA
MICROORGANISMS
MITOCHONDRIA
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PLANT CELLS
RECOMBINANT DNA
RNA
STRUCTURAL CHEMICAL ANALYSIS