scholarly journals Erratum to: A novel thermostable protein-tag: optimization of the Sulfolobus solfataricus DNA-alkyl-transferase by protein engineering

Extremophiles ◽  
2015 ◽  
Vol 20 (1) ◽  
pp. 15-17
Author(s):  
Antonella Vettone ◽  
Mario Serpe ◽  
Aurelio Hidalgo ◽  
José Berenguer ◽  
Giovanni del Monaco ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Sulfolobus Solfataricus ◽  
Protein Tag ◽  
Thermostable Protein

A novel thermostable protein-tag: optimization of the Sulfolobus solfataricus DNA- alkyl-transferase by protein engineering

Extremophiles ◽  
2015 ◽  
Vol 20 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Antonella Vettone ◽  
Mario Serpe ◽  
Aurelio Hidalgo ◽  
José Berenguer ◽  
Giovanni del Monaco ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Sulfolobus Solfataricus ◽  
Protein Tag ◽  
Thermostable Protein

Solution structure and DNA-binding properties of a thermostable protein from the archaeon Sulfolobus solfataricus

1994 ◽  
Vol 1 (11) ◽  
pp. 808-819 ◽  
Author(s):  
Herbert Baumann ◽  
Stefan Knapp ◽  
Thomas Lundbäck ◽  
Rudolf Ladenstein ◽  
Torleif Härd
Keyword(s):  
Dna Binding ◽  
Solution Structure ◽  
Sulfolobus Solfataricus ◽  
Binding Properties ◽  
Thermostable Protein ◽  
Dna Binding Properties

scholarly journals Increased Processivity, Misincorporation, and Nucleotide Incorporation Efficiency in Sulfolobus solfataricus Dpo4 Thumb Domain Mutants

2017 ◽  
Vol 83 (18) ◽  
Author(s):  
Li Wang ◽  
Chenchen Liang ◽  
Jing Wu ◽  
Liming Liu ◽  
Keith E. J. Tyo
Keyword(s):  
Protein Engineering ◽  
Sulfolobus Solfataricus ◽  
Catalytic Efficiency ◽  
Wild Type ◽  
Primer Extension Assay ◽  
Base Pairs ◽  
Content Type ◽  
Thumb Domain ◽  
Nucleotide Incorporation ◽  
Incorporation Efficiency

ABSTRACT The present study aimed to increase the processivity of Sulfolobus solfataricus DNA polymerase Dpo4. Protein engineering and bioinformatics were used to compile a library of potential Dpo4 mutation sites. Ten potential mutants were identified and constructed. A primer extension assay was used to evaluate the processivity of Dpo4 mutants. Thumb (A181D) and finger (E63K) domain mutants showed a processivity of 20 and 19 nucleotides (nt), respectively. A little finger domain mutant (I248Y) exhibited a processivity of 17 nt, only 1 nt more than wild-type Dpo4. Furthermore, the A181D mutant showed lower fidelity and higher nucleotide incorporation efficiency (4.74 × 10−4 s−1 μM−1) than E63K and I248Y mutants. When tasked with bypassing damage, the A181D mutant exhibited a 3.81-fold and 2.62-fold higher catalytic efficiency (k cat/Km ) at incorporating dCTP and dATP, respectively, than wild-type Dpo4. It also showed a 55% and 91.5% higher catalytic efficiency when moving beyond the damaged 8-oxoG:C and 8-oxoG:A base pairs, respectively, compared to wild-type Dpo4. Protein engineering and bioinformatics methods can effectively increase the processivity and translesion synthesis ability of Dpo4. IMPORTANCE DNA polymerases with poor fidelity can be exploited to store data and record changes in response to the intracellular environment. Sulfolobus solfataricus Dpo4 is such an enzyme, although its use is hindered by its low processivity. In this work, we used a bioinformatics and protein engineering approach to generate Dpo4 mutants with improved processivity. We identified the Dpo4 thumb domain as the most relevant in controlling processivity.

Protein engineering on enzymes of the peptide elongation cycle in Sulfolobus solfataricus

Biochimie ◽  
1998 ◽  
Vol 80 (11) ◽  
pp. 895-898 ◽  
Author(s):  
Vincenzo Bocchini ◽  
Bianca Stella Adinolfi ◽  
Paolo Arcari ◽  
Alessandro Arcucci ◽  
Antonio Dello Russo ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Sulfolobus Solfataricus ◽  
Elongation Cycle ◽  
Peptide Elongation

Structure and stability of a thioredoxin reductase from Sulfolobus solfataricus: A thermostable protein with two functions

2009 ◽  
Vol 1794 (3) ◽  
pp. 554-562 ◽  
Author(s):  
Alessia Ruggiero ◽  
Mariorosario Masullo ◽  
Maria Rosaria Ruocco ◽  
Pasquale Grimaldi ◽  
Maria Angela Lanzotti ◽  
...  
Keyword(s):  
Sulfolobus Solfataricus ◽  
Thioredoxin Reductase ◽  
Structure And Stability ◽  
Thermostable Protein

Insights into the biochemical and functional characterization of sortase E transpeptidase of Corynebacterium glutamicum

2019 ◽  
Vol 476 (24) ◽  
pp. 3835-3847 ◽  
Author(s):  
Aliyath Susmitha ◽  
Kesavan Madhavan Nampoothiri ◽  
Harsha Bajaj
Keyword(s):  
Protein Engineering ◽  
Cell Attachment ◽  
Functional Characterization ◽  
Protein Structures ◽  
Structural Similarity ◽  
Surface Proteins ◽  
Sortase A ◽  
Peptide Cleavage ◽  
New Findings

Most Gram-positive bacteria contain a membrane-bound transpeptidase known as sortase which covalently incorporates the surface proteins on to the cell wall. The sortase-displayed protein structures are involved in cell attachment, nutrient uptake and aerial hyphae formation. Among the six classes of sortase (A–F), sortase A of S. aureus is the well-characterized housekeeping enzyme considered as an ideal drug target and a valuable biochemical reagent for protein engineering. Similar to SrtA, class E sortase in GC rich bacteria plays a housekeeping role which is not studied extensively. However, C. glutamicum ATCC 13032, an industrially important organism known for amino acid production, carries a single putative sortase (NCgl2838) gene but neither in vitro peptide cleavage activity nor biochemical characterizations have been investigated. Here, we identified that the gene is having a sortase activity and analyzed its structural similarity with Cd-SrtF. The purified enzyme showed a greater affinity toward LAXTG substrate with a calculated KM of 12 ± 1 µM, one of the highest affinities reported for this class of enzyme. Moreover, site-directed mutation studies were carried to ascertain the structure functional relationship of Cg-SrtE and all these are new findings which will enable us to perceive exciting protein engineering applications with this class of enzyme from a non-pathogenic microbe.

Protein Engineering of Penicillin Acylase

Acta Naturae ◽  
2010 ◽  
Vol 2 (3) ◽  
pp. 47-61 ◽  
Author(s):  
V I Tishkov ◽  
S S Savin ◽  
A S Yasnaya
Keyword(s):  
Protein Engineering ◽  
Penicillin Acylase

Exemplar Abstract for Sulfolobus solfataricus Zillig et al. 1980.

2003 ◽  
Author(s):  
Charles Thomas Parker ◽  
Dorothea Taylor ◽  
George M Garrity
Keyword(s):  
Sulfolobus Solfataricus

Self-Assembling Micelles Based on an Intrinsically Disordered Protein Domain

2018 ◽  
Author(s):  
Sarah Klass ◽  
Matthew J. Smith ◽  
Tahoe Fiala ◽  
Jessica Lee ◽  
Anthony Omole ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Fusion Proteins ◽  
Organic Molecules ◽  
Intrinsically Disordered Protein ◽  
Protein Domain ◽  
Enzymatic Cleavage ◽  
Intrinsically Disordered ◽  
Disordered Protein ◽  
Self Assembling ◽  
Protein Tag

Herein, we describe a new series of fusion proteins that have been developed to self-assemble spontaneously into stable micelles that are 27 nm in diameter after enzymatic cleavage of a solubilizing protein tag. The sequences of the proteins are based on a human intrinsically disordered protein, which has been appended with a hydrophobic segment. The micelles were found to form across a broad range of pH, ionic strength, and temperature conditions, with critical micelle concentration (CMC) values below 1 µM being observed in some cases. The reported micelles were found to solubilize hydrophobic metal complexes and organic molecules, suggesting their potential suitability for catalysis and drug delivery applications.

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