Characterization of a new hemoprotein in the yeast Saccharomyces cerevisiae.
The Saccharomyces cerevisiae gene YNL234w encodes a 426-amino acid-long protein that shares significant similarities with the globin family. Compared with known globins from unicellular organisms, the Ynl234wp polypeptide is characterized by an unusual structure. In this protein, a central putative heme-binding domain of about 140 amino acids is flanked by two sequences of about 160 and 120 amino acids, respectively, which share no similarity with known polypeptides. Northern analysis indicates that YNL234w transcription is very low in cells grown under normal aerobic conditions but is induced by oxygen-limited growth conditions and by other stress conditions such as glucose repression, heat shock, osmotic stress, and nitrogen starvation. However, the deletion of the gene had no detectable effect on yeast growth. The Ynl234wp polypeptide has been expressed in Escherichia coli, and the hemoprotein nature of the recombinant protein was demonstrated by heme staining after SDS/polyacrylamide gel electrophoresis and spectroscopic analysis. Our data indicate that purified recombinant Ynl234wp possesses a noncovalently bound heme molecule that is predominantly found in a low spin form.
Sartori
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Dipartimento di Chimica Biologica, viale G. Colombo, 3, 35121 Padova, Italy.
Aldegheri
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Mazzotta
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Lanfranchi
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Tournu
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Brown
A J
AJ
Carignani
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D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D006420:Hemeproteins / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic