Primary structure of spinach-chloroplast thioredoxin f. Protein sequencing and analysis of complete cDNA clones for spinach-chloroplast thioredoxin f.
The primary structure of thioredoxin f from spinach chloroplasts was determined by standard amino acid sequencing and furthermore by sequencing the corresponding nuclear genome region. The protein, with a calculated molecular mass of 12,564 Da and a molar absorption coefficient at 280 nm of 17,700 M-1 cm-1, consists of 113 residues and exhibits 24% residue identities with spinach chloroplast thioredoxin mb or Escherichia coli thioredoxin. A monospecific antibody elicited against thioredoxin f has been used to select recombinant phage from spinach cDNA libraries in lambda gt11. The inserts of positive clones were sequenced. They code for a polypeptide of 190 amino acids, composed of the thioredoxin f sequence (113 residues) and an upstream element (77 residues) which most probably constitutes the N-terminal transit peptide that directs the polypeptide into chloroplasts. In vitro transcription and translation of this construct generates a polypeptide of approximately 21 kDa, which is imported by isolated spinach chloroplasts and processed to the mature 12.5-kDa protein.
Kamo
M
M
Research Institute for Biosciences, Science University of Tokyo.
Tsugita
A
A
Wiessner
C
C
Wedel
N
N
Bartling
D
D
Herrmann
R G
RG
Aguilar
F
F
Gardet-Salvi
L
L
Schürmann
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D054479:Chloroplast Thioredoxins; D002736:Chloroplasts / Q000032:analysis*; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D010945:Plants, Edible / Q000032:analysis*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013879:Thioredoxins; D014158:Transcription, Genetic
D054479:Chloroplast Thioredoxins; D010446:Peptide Fragments; D010940:Plant Proteins; D011994:Recombinant Proteins; D013879:Thioredoxins; D004247:DNA