Nitrite reductase from Pseudomonas aeruginosa: Sequence of the gene and the protein

Abstract

The gene coding for nitrite reductase of Pseudomonas aeruginosa has been cloned and its sequence determined. The coding region is 1707 bp long and contains information for a polypeptide chain of 568 amino acids. The sequence of the mature protein has been confirmed independently by extensive amino acid sequencing. The amino-terminus of the mature protein is located at Lys-26; the preceding 25 residue long extension shows the features typical of signal peptides. Therefore the enzyme is probably secreted into the periplasmic space. The mature protein is made of 543 amino acid residues and has a molecular mass of 60204 Da. The c-heme-binding domain, which contains the only two Cys of the molecule, is located at the amino-terminal region. Analysis of the protein sequence in terms of hydrophobicity profile gives results consistent with the fact that the enzyme is fully water soluble and not membrane bound; the most hydrophilic region appears to correspond to the c-heme domain. Secondary structure predictions are in general agreement with previous analysis of circular dichroic data.Nitrite reductase; Primary structure; Pre-protein; Cytochrome oxidase; (Pseudomonas aeruginosa)

The structural gene for cytochrome c551 from Pseudomonas aeruginosa

“…The first is the C-terminal of nitrite reductase [5] and the second is cytoehrome ~551. Analysis of the nucleotide sequence shows that cytochrome ~551 is expressed as a 104 amino acid preprotein.…”

“…Cytochrome ~551 is located in the periplasmic space of Pseudomonas aeruginosa [2], and delivers electrons to nitrite reductase (EC 1.9.3.2) either directly or via azurin [2,3]. These 3 proteins in the Pseudomonas respiratory chain are located in the periplasmic space [2]; this is also confirmed by the nucleotide sequence in the 3 genes, showing the existence of a signal peptide [4,5].…”

“…This is in accordance with the findings of Wood [2] who reports an almost 1: 1 stoichiometry between cytochrome ~551 and the nitrite reductase monomer in Pseudomonas aeruginosa grown under anaerobic conditions. The tr~scriptional terminator suggested by Silvestrini [5] may then act as a transcriptional pause signal. Cloning the promoter region of the operon will give valuable information about the regulation.…”

Regulation of oxidative phosphorylation: The flexible respiratory network ofParacoccus denitrificans

“…The nirS gene encoding cd~ type nitrate reductase has been cloned from Pseudomonas stutzeri (Smith and Tiedje, 1992), Ps. aeruginosa (Silvestrini et al, 1989), and recently from P. denitrificans . The P. denitrificans nirS gene is located in a gene cluster (Fig.…”

Transcriptional analysis of the nirS gene, encoding cytochrome cd 1 nitrite reductase, of Paracoccus pantotrophus LMD 92.63 The GenBank accession number for the sequence in this paper is U75413.

“…The latter haem has only been found in denitrifiers containing this type of nitrite reductase. Di-haem cd " nitrite reductases have been (Moir et al, 1993 ;Baker et al, 1997) as well as in other denitrifiers including Pseudomonas aeruginosa (Silvestrini et al, 1989), Pseudomonas stutzeri Zobell (Ju$ ngst et al, 1991), Ps. stutzeri JM300 (Smith & Tiedje, 1992), Paracoccus denitrificans IFO 12442 (Ohshima et al, 1993), Pa. denitrificans PD1222 (de Boer et al, 1994) and Ralstonia eutropha H16 (Rees et al, 1997).…”