Proteome analysis of an aerobic hyperthermophilic crenarchaeon, Aeropyrum pernix K1.
We analyzed the proteome of a crenararchaeon, Aeropyrum pernix K1, by using the following four methods: (i) two-dimensional PAGE followed by MALDI-TOF MS, (ii) one-dimensional SDS-PAGE in combination with two-dimensional LC-MS/MS, (iii) multidimensional LC-MS/MS, and (iv) two-dimensional PAGE followed by amino-terminal amino acid sequencing. These methods were found to be complementary to each other, and biases in the data obtained in one method could largely be compensated by the data obtained in the other methods. Consequently a total of 704 proteins were successfully identified, 134 of which were unique to A. pernix K1, and 19 were not described previously in the genomic annotation. We found that the original annotation of the genomic data of this archaeon was not adequate in particular with respect to proteins of 10-20 kDa in size, many of which were described as hypothetical. Furthermore the amino-terminal amino acid sequence analysis indicated that surprisingly the translation of 52% of their genes starts with TTG in contrast to ATG (28%) and GTG (20%). Thus, A. pernix K1 is the first example of an organism in which TTG is the most predominant translational initiation codon.
Yamazaki
Syuji
S
Department of Biotechnology, Genome Analysis Division, National Institute of Technology and Evaluation, Shibuya, Tokyo 151-0066, Japan. yamazaki-shuji@nite.go.jp
Yamazaki
Jun
J
Nishijima
Keiko
K
Otsuka
Rie
R
Mise
Miyako
M
Ishikawa
Hanako
H
Sasaki
Kazumi
K
Tago
Shin-ichi
S
Isono
Katsumi
K
D000332:Aerobiosis; D041522:Aeropyrum / Q000737:chemistry* / Q000145:classification; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry; D001482:Base Composition / Q000235:genetics; D002853:Chromatography, Liquid; D003062:Codon / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D057927:Hydrophobic and Hydrophilic Interactions; D007526:Isoelectric Point; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D010442:Peptide Chain Initiation, Translational / Q000235:genetics; D020543:Proteome / Q000032:analysis* / Q000737:chemistry*; D040901:Proteomics
D019843:Archaeal Proteins; D003062:Codon; D020543:Proteome