Glutamine deamidase
Monday, May 1, 2023
8:18 PM
Glutamine deamidase General information: 1. Gene 10 in the Lewando phage 2. Gene start site: 5769, Gene stop site: 6860 3. Gene length: 1091 bp 4. Direction: Forward 5. Gene function: glutamine deamidase 6. Protein sequence: MPSVSDEVVDDFLAHYGVKGMRWGHRKSEEVSPERQAKREERAQKFERKAQM LDTKISEIANASHLKDWDKAYQIQELEAHKRTALSDAEAKRQGKLSAKQKKVIK GAAIAAAIVAAYLIQDNVQSGEFTRLIAKGKAHFGEEDFKFKKNSDLLNPHFTAD DIHEKVVKHVNPDYGGIGTKMNCRRATFSYEMRRRGYDVKATKTTNGNGQHVL GLHNAITPDEKFKSTHRLNVTKTAISQVVSKAKGKDAPILDLTENFAAGGRNKIE NGSSPFGIFDALSKQPDGSRGELGVMWSMGGGHSMAYEVFNGKAIVFDAQSGK KLDTPEKFKEAMGSVDTAGFTRLDNVELNHDYLMRWMTNVKZ 7. Nucleotide sequence: ATGCCTTCCGTGAGTGATGAAGTAGTAGACGATTTCCTTGCTCACTATGGCGTA AAGGGTATGCGATGGGGCCATCGTAAATCTGAAGAGGTTAGTCCTGAGCGGCA GGCTAAGCGGGAAGAGCGTGCTCAGAAGTTTGAGCGTAAAGCTCAGATGCTG GATACGAAAATCAGTGAGATCGCAAATGCTTCCCATTTGAAAGATTGGGATAA GGCATATCAGATCCAAGAACTGGAAGCGCATAAGCGCACAGCTCTCTCCGATG CCGAAGCCAAGAGGCAGGGCAAGCTCTCCGCCAAGCAGAAAAAGGTCATTA AGGGTGCAGCTATTGCCGCCGCTATCGTGGCGGCATATCTCATTCAGGACAATG TTCAGAGTGGCGAGTTCACTCGTCTGATAGCTAAAGGTAAAGCCCACTTTGGC GAAGAGGATTTCAAGTTCAAGAAGAACTCGGATCTGTTGAATCCCCACTTCAC TGCTGACGATATCCACGAGAAGGTAGTCAAGCACGTCAATCCTGACTACGGTG GAATCGGAACCAAGATGAACTGTCGTCGTGCAACATTCTCATATGAGATGCGT CGTCGAGGATATGACGTCAAGGCAACTAAGACCACTAATGGTAATGGTCAGCA TGTTTTGGGTCTGCACAATGCAATCACTCCTGATGAGAAGTTCAAAAGCACAC ATCGACTCAACGTCACTAAGACTGCGATTTCGCAAGTCGTCAGTAAGGCAAAA GGTAAGGATGCACCTATCCTCGACCTGACTGAGAACTTTGCAGCAGGTGGTCG TAACAAGATCGAGAATGGCAGTAGCCCTTTCGGTATATTCGATGCCCTATCCAA ACAGCCCGACGGTTCACGCGGTGAGCTCGGTGTGATGTGGAGTATGGGTGGC GGGCACAGTATGGCGTATGAAGTCTTCAACGGTAAGGCTATCGTTTTCGATGCT CAGTCCGGCAAGAAGCTCGATACACCTGAGAAGTTCAAAGAAGCAATGGGGT CTGTCGACACCGCTGGCTTTACCCGACTGGACAACGTTGAGTTGAACCACGAT TACCTCATGAGGTGGATGACCAATGTTAAGTGA Evidence: 1. Blaspt on DNA master showed high alignment with the Sporto gene 8 a. Alignment of 98.9% b. E-value of 0.0e0 c. 9:5 alignment (query: target) 2. HHpred showed high probability with glutamine deamidase a. Papain fold toxin 1, glutamine deamidase i. 98.47% probability ii. Score of 66.48 iii. Target length of 112 iv. [link] b. ORF169b; Shigella Bacterial protein i. 96.61% probability ii. Score of 50.78 iii. Target length of 220 iv. [link] 3. Literature: a. The Shigella flexneri effector OspI deamidates UBC13 to dampen the inflammatory response i. Sanada, T., Kim, M., Mimuro, H., Suzuki, M., Ogawa, M., Oyama, A., Ashida, H., Kobayashi, T., Koyama, T., Nagai, S., Shibata, Y., Gohda, J., Inoue, J.I., Mizushima, T., Sasakawa, C. ii. (2012) Nature 483: 623-626 iii. DOI: [link] 4. Catalytic triad of of Shigella flexneri Ospl glutamine deamidase protein found in the lewando gene 10 a. Catalytic triad: C62, H145, D160 b. Figure 1 5. Pfam glutamine deamidase signature a. Lewando gene 10 also contains conserved nucleotides found in the glutamine deamidase signature b. Figure 2, 3 c. [link] Figure 1: Identification of the catalytic triad conserved in the glutamine deamidase. Q_4214840 corresponds to the protein sequence of gene #10 in the Lewando phage. T 3B21_A corresponds to the sequence found in the Ospl protein. The Blue rectangles illustrate the location of the catalytic triad conserved in glutamine deamidases. Both sequences contained the necessary CHD catalytic triad. Figure 2: Glutamine deamidase signature. Glutamine deamidase signature shows the most conserved nucleotides when comparing multiple glutamine damidases. Within this signature is the catalytic triad (C62, H145, D160) along with other conserved nucleotides. Figure 3: Locating conserved nucleotides found in glutamine deamidase signature. Clustal omega was used to align the sequences of the Ospl Shigella protein (3B21_1), Lewando gene 10 (Lewando_10), and Sporto gene 8 (Sporto_8). Boxed in red are the conserved nucleotides found in the glutamine deamidase signature.