scholarly journals ANTICALIgN: visualizing, editing and analyzing combined nucleotide and amino acid sequence alignments for combinatorial protein engineering

2016 ◽  
Vol 29 (7) ◽  
pp. 263-270 ◽  
Author(s):  
Alexander Jarasch ◽  
Melanie Kopp ◽  
Evelyn Eggenstein ◽  
Antonia Richter ◽  
Michaela Gebauer ◽  
...  
Keyword(s):  
Amino Acid ◽  
Protein Engineering ◽  
Amino Acid Sequence ◽  
Sequence Alignments ◽  
Combinatorial Protein Engineering

scholarly journals Broad-spectrum detection of papillomaviruses in bovine teat papillomas and healthy teat skin

2004 ◽  
Vol 85 (8) ◽  
pp. 2191-2197 ◽  
Author(s):  
Tomoko Ogawa ◽  
Yoshimi Tomita ◽  
Mineyuki Okada ◽  
Kuniko Shinozaki ◽  
Hiroko Kubonoya ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Broad Spectrum ◽  
Phylogenetic Analyses ◽  
Genomic Diversity ◽  
Bovine Papillomavirus ◽  
Subclinical Infection ◽  
Healthy Skin ◽  
Sequence Alignments ◽  
Spectrum Detection

To investigate the prevalence of bovine papillomavirus (BPV) in bovine papilloma and healthy skin, DNA extracted from teat papillomas and healthy teat skin swabs was analysed by PCR using the primer pairs FAP59/FAP64 and MY09/MY11. Papillomavirus (PV) DNA was detected in all 15 papilloma specimens using FAP59/FAP64 and in 8 of the 15 papilloma specimens using MY09/MY11. In swab samples, 21 and 8 of the 122 samples were PV DNA positive using FAP59/FAP64 and MY09/MY11, respectively. Four BPV types (BPV-1, -3, -5 and -6), two previously identified putative BPV types (BAA1 and -5) and 11 putative new PV types (designated BAPV1 to -10 and BAPV11MY) were found in the 39 PV DNA-positive samples. Amino acid sequence alignments of the putative new PV types with reported BPVs and phylogenetic analyses of the putative new PV types with human and animal PV types showed that BAPV1 to -10 and BAPV11MY are putative new BPV types. These results also showed the genomic diversity and extent of subclinical infection of BPV.

scholarly journals A Novel Lipolytic Enzyme Located in the Outer Membrane of Pseudomonas aeruginosa

1999 ◽  
Vol 181 (22) ◽  
pp. 6977-6986 ◽  
Author(s):  
Susanne Wilhelm ◽  
Jan Tommassen ◽  
Karl-Erich Jaeger
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Outer Membrane ◽  
Open Reading Frame ◽  
Lipolytic Enzyme ◽  
Cell Fractionation ◽  
Sequence Alignments ◽  
Reading Frame ◽  
Amino Terminal

ABSTRACT A lipase-negative deletion mutant of Pseudomonas aeruginosa PAO1 still showed extracellular lipolytic activity toward short-chain p-nitrophenylesters. By screening a genomic DNA library of P. aeruginosa PAO1, an esterase gene, estA, was identified, cloned, and sequenced, revealing an open reading frame of 1,941 bp. The product ofestA is a 69.5-kDa protein, which is probably processed by removal of an N-terminal signal peptide to yield a 67-kDa mature protein. A molecular mass of 66 kDa was determined for35S-labeled EstA by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The amino acid sequence of EstA indicated that the esterase is a member of a novel GDSL family of lipolytic enzymes. The estA gene showed high similarity to an open reading frame of unknown function located in thetrpE-trpG region of P. putida and to a gene encoding an outer membrane esterase of Salmonella typhimurium. Amino acid sequence alignments led us to predict that this esterase is an autotransporter protein which possesses a carboxy-terminal β-barrel domain, allowing the secretion of the amino-terminal passenger domain harboring the catalytic activity. Expression of estA in P. aeruginosa andEscherichia coli and subsequent cell fractionation revealed that the enzyme was associated with the cellular membranes. Trypsin treatment of whole cells released a significant amount of esterase, indicating that the enzyme was located in the outer membrane with the catalytic domain exposed to the surface. To our knowledge, this esterase is unique in that it exemplifies in P. aeruginosa(i) the first enzyme identified in the outer membrane and (ii) the first example of a type IV secretion mechanism.

scholarly journals SBAL: a practical tool to generate and edit structure-based amino acid sequence alignments

2012 ◽  
Vol 28 (7) ◽  
pp. 1026-1027 ◽  
Author(s):  
Conan K. Wang ◽  
Ursula Broder ◽  
Saroja K. Weeratunga ◽  
Robin B. Gasser ◽  
Alex Loukas ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Sequence Alignments ◽  
Practical Tool

scholarly journals The Metaxin Mitochondrial Import Proteins: Multiple Metaxin-like Proteins in Fungi

2022 ◽  
Author(s):  
Kenneth W Adolph
Keyword(s):  
Phylogenetic Analysis ◽  
Amino Acid ◽  
Membrane Proteins ◽  
Amino Acid Sequence ◽  
Mitochondrial Membrane ◽  
Protein Domains ◽  
Secondary Structures ◽  
Sequence Alignments ◽  
Mitochondrial Import ◽  
High Degree

Multiple metaxin-like proteins are shown to exist in fungi, as also found for the metaxin proteins of vertebrates and invertebrates. In vertebrates, metaxins 1 and 2 are mitochondrial membrane proteins that function in the import of proteins into mitochondria. Fungal metaxin-like proteins were identified by criteria including their homology with human metaxins and the presence of characteristic GST_N_Metaxin, GST_C_Metaxin, and Tom37 protein domains. Fungi in different taxonomic divisions (phyla) were found to possess multiple metaxin-like proteins. These include the Ascomycota, Basidiomycota, Blastocladiomycota, Chytridiomycota, Mucoromycota, Neocallimastigomycota, and Zoopagomycota divisions. Most fungi with multiple metaxin-like proteins contain two proteins, designated MTXa and MTXb. Amino acid sequence alignments show a high degree of homology among MTXa proteins, with over 60% amino acid identities, and also among MTXb proteins of fungi in the same division. But very little homology is observed in aligning MTXa with MTXb proteins of the same or different fungi. Both the MTXa proteins and MTXb proteins have the protein domains that characterize the metaxins and metaxin-like proteins: GST_N_Metaxin, GST_C_Metaxin, and Tom37. The metaxins and metaxin-like proteins of vertebrates, invertebrates, plants, protists, and bacteria all possess these domains. The secondary structures of MTXa and MTXb proteins are both dominated by similar patterns of α-helical segments, but extensive β-strand segments are absent. Nine highly conserved α-helical segments are present, the same as other metaxins and metaxin-like proteins. Phylogenetic analysis reveals that MTXa and MTXb proteins of fungi form two separate and distinct groups. These groups are also separate from the groups of vertebrate metaxins, metaxin-related Sam37 proteins of yeasts, and metaxin-like FAXC proteins.

scholarly journals Apical targeting of the P2Y4 receptor is directed by hydrophobic and basic residues in the cytoplasmic tail

2013 ◽  
Vol 304 (3) ◽  
pp. C228-C239 ◽  
Author(s):  
D. Ross DuBose ◽  
Samuel C. Wolff ◽  
Ai-Dong Qi ◽  
Izabela Naruszewicz ◽  
Robert A. Nicholas
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Basolateral Membrane ◽  
Chloride Secretion ◽  
Common Mechanism ◽  
Main Body ◽  
Sequence Alignments ◽  
Hydrophobic Residues ◽  
Intestinal Chloride Secretion ◽  
Apical Sorting

The P2Y4 receptor is selectively targeted to the apical membrane in polarized epithelial cell lines and has been shown to play a key role in intestinal chloride secretion. In this study, we delimit a 23 amino acid sequence within the P2Y4 receptor C-tail that directs its apical targeting. Using a mutagenesis approach, we found that four hydrophobic residues near the COOH-terminal end of the signal are necessary for apical sorting, whereas two basic residues near the NH2-terminal end of the signal are involved to a lesser extent. Interestingly, mutation of the key hydrophobic residues results in a basolateral enrichment of the receptor construct, suggesting that the apical targeting sequence may prevent insertion or disrupt stability of the receptor at the basolateral membrane. The signal is not sequence specific, as an inversion of the 23 amino acid sequence does not disrupt apical targeting. We also show that the apical targeting sequence is an autonomous signal and is capable of redistributing the normally basolateral P2Y12 receptor, suggesting that the apical signal is dominant over the basolateral signal in the main body of the P2Y12 receptor. The targeting sequence is unique to the P2Y4 receptor, and sequence alignments of the COOH-terminal tail of mammalian orthologs reveal that the hydrophobic residues in the targeting signal are highly conserved. These data define the novel apical sorting signal of the P2Y4 receptor, which may represent a common mechanism for trafficking of epithelial transmembrane proteins.

scholarly journals Defining Electron Bifurcation in the Electron-Transferring Flavoprotein Family

2017 ◽  
Vol 199 (21) ◽  
Author(s):  
Amaya M. Garcia Costas ◽  
Saroj Poudel ◽  
Anne-Frances Miller ◽  
Gerrit J. Schut ◽  
Rhesa N. Ledbetter ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Energy Conservation ◽  
Structural Modeling ◽  
Sequence Motifs ◽  
Quinone Oxidoreductase ◽  
Sequence Alignments ◽  
Electron Transfer Mechanism ◽  
Electron Transferring Flavoprotein

ABSTRACT Electron bifurcation is the coupling of exergonic and endergonic redox reactions to simultaneously generate (or utilize) low- and high-potential electrons. It is the third recognized form of energy conservation in biology and was recently described for select electron-transferring flavoproteins (Etfs). Etfs are flavin-containing heterodimers best known for donating electrons derived from fatty acid and amino acid oxidation to an electron transfer respiratory chain via Etf-quinone oxidoreductase. Canonical examples contain a flavin adenine dinucleotide (FAD) that is involved in electron transfer, as well as a non-redox-active AMP. However, Etfs demonstrated to bifurcate electrons contain a second FAD in place of the AMP. To expand our understanding of the functional variety and metabolic significance of Etfs and to identify amino acid sequence motifs that potentially enable electron bifurcation, we compiled 1,314 Etf protein sequences from genome sequence databases and subjected them to informatic and structural analyses. Etfs were identified in diverse archaea and bacteria, and they clustered into five distinct well-supported groups, based on their amino acid sequences. Gene neighborhood analyses indicated that these Etf group designations largely correspond to putative differences in functionality. Etfs with the demonstrated ability to bifurcate were found to form one group, suggesting that distinct conserved amino acid sequence motifs enable this capability. Indeed, structural modeling and sequence alignments revealed that identifying residues occur in the NADH- and FAD-binding regions of bifurcating Etfs. Collectively, a new classification scheme for Etf proteins that delineates putative bifurcating versus nonbifurcating members is presented and suggests that Etf-mediated bifurcation is associated with surprisingly diverse enzymes. IMPORTANCE Electron bifurcation has recently been recognized as an electron transfer mechanism used by microorganisms to maximize energy conservation. Bifurcating enzymes couple thermodynamically unfavorable reactions with thermodynamically favorable reactions in an overall spontaneous process. Here we show that the electron-transferring flavoprotein (Etf) enzyme family exhibits far greater diversity than previously recognized, and we provide a phylogenetic analysis that clearly delineates bifurcating versus nonbifurcating members of this family. Structural modeling of proteins within these groups reveals key differences between the bifurcating and nonbifurcating Etfs.

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