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

Journal Article · Fri Mar 15 00:00:00 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), Vol. 88:6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

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