Practice 7. Uniprot

This research will examine the protein bifunctional phosphoglucose/phosphomannose isomerase of the archaea Pyrobaculum aerophilum

Keywords: Pyrobaculum aerophilum, enzyme

Function:

Catalyzes isomerization of glucose-6-phosphate and mannose-6-phosphate with equal catalytic efficiency.

About the organism

Pyrobaculum aerophilum - facultatively aerobic nitrate-reducing hyperthermophilic archaea. This organism has a number of interesting features. For example, computational analysis of the genome has confirmed experiments showing that P. aerophilum (and possibly all crenarchaea) do not have 5' untranslated regions of mRNA and thus do not appear to use a mechanism based on the ribosome binding site to initiate translation at the 5'-end.[1] Pyrobaculum aerophilum got its name from the Greek noun "aer" (air) and the Greek adjective "philos" (loving). It was found that optimal growth conditions are achieved at 100°C and pH 7.0[2].

Commands used to download proteomes:

wget 'https://www.uniprot.org/uniprot/?query=proteome:up000002439&format=txt&compress=yes' -O UP000002439.swiss.gz

wget 'https://www.uniprot.org/uniprot/?query=proteome:up000001694&format=txt&compress=yes' -O UP000001694.swiss.gz

The reference proteome of archaea Pyrobaculum aerophilum was chosen for comparison, Thermoproteus neutrophilus proteome was taken as a control. (a related strain that is capable of producing hydrogen sulfide)..

Pyrobaculum aerophilum

ID:UP00002439

Number of proteins in the proteome: 2590 (в swiss-prot:315). Described 12,16%

BUSCO: C:96.5%[S:96.3%,D:0.2%],F:0.2%,M:3.2%,n:404 thermoproteales_odb10. Therefore, 96.5% complete.

CPD is close to standard (slightly higher)

Thermoproteus neutrophilus

ID: UP000001694

Number of proteins in the proteome: 1965 (в swiss-prot:148). Described 7,53%

BUSCO: C:95.8%[S:95.5%,D:0.2%],F:0.2%,M:4%,n:404 thermoproteales_odb10. Therefore, 95.8% complete.

CPD standard

Pyrobaculum aerophilum

Command to search for transmembrane proteins:

annotation:(type:transmem) AND organism:"Pyrobaculum aerophilum (strain ATCC 51768 / DSM 7523 / JCM 9630 / CIP 104966 / NBRC 100827 / IM2) [178306]" AND proteome:up000002439

Number of transmembrane proteins: 475 (fraction in proteome: 18,34%)

Enzyme search command:

ec:* AND organism:"Pyrobaculum aerophilum (strain ATCC 51768 / DSM 7523 / JCM 9630 / CIP 104966 / NBRC 100827 / IM2) [178306]" AND proteome:up000002439

Number of enzymes:336 (fraction in proteome:12,97%)

Command to search for oxidoreductases:

ec:1* AND organism:"Pyrobaculum aerophilum (strain ATCC 51768 / DSM 7523 / JCM 9630 / CIP 104966 / NBRC 100827 / IM2) [178306]" AND proteome:up000002439

Number of oxidoreductases: 48 (fraction in proteome:1,85%)

Thermoproteus neutrophilus

Command to search for transmembrane proteins:

annotation:(type:transmem) AND organism:"Pyrobaculum neutrophilum (strain DSM 2338 / JCM 9278 / NBRC 100436 / V24Sta) (Thermoproteus neutrophilus) [444157]" AND proteome:up000001694

Number of transmembrane proteins:318 (fraction in proteome:16,18%)

Enzyme search command:

ec:* AND organism:"Pyrobaculum neutrophilum (strain DSM 2338 / JCM 9278 / NBRC 100436 / V24Sta) (Thermoproteus neutrophilus) [444157]" AND proteome:up000001694

Number of enzymes:336 (fraction in proteome:17%)

Command to search for oxidoreductases:

ec:1* AND organism:"Pyrobaculum neutrophilum (strain DSM 2338 / JCM 9278 / NBRC 100436 / V24Sta) (Thermoproteus neutrophilus) [444157]" AND proteome:up000001694

Number of oxidoreductases: 51(2,6%)

From the above data, we can see that the proportion of oxidoreductases in Thermoproteus neutrophilus, which is capable of forming hydrogen sulfide,is still greater than in Pyrobaculum aerophilum (H2S synthesis involves oxidoreductases). We can assume that the percentage of oxidoreductases does not differ significantly because Pyrobaculum aerophilum uses the mechanism of nitrogen reduction in the process of life activity..

Comparison of proteomes by first amino acid:

Commands to check the first amino acid:

zcat UP000002439.swiss | seqret -filter 'swiss::stdin:*[1:1]' | grep -v "^>" | uniq

zcat UP000001694.swiss | seqret -filter 'swiss::stdin:*[1:1]' | grep -v "^>" | uniq

As a result of running the program, we were able to establish that in both cases all proteins start with methionine (M). This is due to the fact that the AUG triplet is a start codon and encodes methionine. It should be noted that in prokaryotes it is read as formylmethionine..

Sources

1. Genome sequence of the hyperthermophilic crenarchaeon Pyrobaculum aerophilum Sorel T Fitz-Gibbon 1, Heidi Ladner, Ung-Jin Kim, Karl O Stetter, Melvin I Simon, Jeffrey H Miller

2. P Völkl, R Huber, E Drobner, R Rachel, S Burggraf, A Trincone, and K O Stetter. "Pyrobaculum aerophilum sp. nov., a Novel Nitrate-Reducing Hyperthermophilic Archaeum." Applied and Environmental Microbiology, Sept. 1993, p. 2918-2926; 0099-2240/93/092918.