Design of a Fe 4 S 4 cluster into the core of a de novo four‐helix bundle

2020 ◽  
Vol 67 (4) ◽  
pp. 574-585 ◽  
Author(s):  
Joshua A. Mancini ◽  
Douglas H. Pike ◽  
Alexei M. Tyryshkin ◽  
Liti Haramaty ◽  
Michael S. Wang ◽  
...  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shin Irumagawa ◽  
Kaito Kobayashi ◽  
Yutaka Saito ◽  
Takeshi Miyata ◽  
Mitsuo Umetsu ◽  
...  

AbstractThe stability of proteins is an important factor for industrial and medical applications. Improving protein stability is one of the main subjects in protein engineering. In a previous study, we improved the stability of a four-helix bundle dimeric de novo protein (WA20) by five mutations. The stabilised mutant (H26L/G28S/N34L/V71L/E78L, SUWA) showed an extremely high denaturation midpoint temperature (Tm). Although SUWA is a remarkably hyperstable protein, in protein design and engineering, it is an attractive challenge to rationally explore more stable mutants. In this study, we predicted stabilising mutations of WA20 by in silico saturation mutagenesis and molecular dynamics simulation, and experimentally confirmed three stabilising mutations of WA20 (N22A, N22E, and H86K). The stability of a double mutant (N22A/H86K, rationally optimised WA20, ROWA) was greatly improved compared with WA20 (ΔTm = 10.6 °C). The model structures suggested that N22A enhances the stability of the α-helices and N22E and H86K contribute to salt-bridge formation for protein stabilisation. These mutations were also added to SUWA and improved its Tm. Remarkably, the most stable mutant of SUWA (N22E/H86K, rationally optimised SUWA, ROSA) showed the highest Tm (129.0 °C). These new thermostable mutants will be useful as a component of protein nanobuilding blocks to construct supramolecular protein complexes.


2012 ◽  
Vol 18 (50) ◽  
pp. 15890-15890
Author(s):  
Marina Faiella ◽  
Ornella Maglio ◽  
Flavia Nastri ◽  
Angela Lombardi ◽  
Liliana Lista ◽  
...  

1998 ◽  
Vol 120 (40) ◽  
pp. 10287-10295 ◽  
Author(s):  
Kerstin S. Broo ◽  
Helena Nilsson ◽  
Jonas Nilsson ◽  
Lars Baltzer

2015 ◽  
Vol 2015 (21) ◽  
pp. 3352-3352 ◽  
Author(s):  
Marco Chino ◽  
Ornella Maglio ◽  
Flavia Nastri ◽  
Vincenzo Pavone ◽  
William F. DeGrado ◽  
...  

Science ◽  
1990 ◽  
Vol 249 (4971) ◽  
pp. 884-891 ◽  
Author(s):  
M. Hecht ◽  
J. Richardson ◽  
D. Richardson ◽  
R. Ogden

2008 ◽  
Vol 130 (29) ◽  
pp. 9188-9189 ◽  
Author(s):  
Jennifer R. Calhoun ◽  
Caleb B. Bell ◽  
Thomas J. Smith ◽  
Thomas J. Thamann ◽  
William F. DeGrado ◽  
...  

Author(s):  
Angela Lombardi ◽  
Fabio Pirro ◽  
Ornella Maglio ◽  
Marco Chino ◽  
William F. DeGrado

2006 ◽  
Vol 128 (2) ◽  
pp. 663-663 ◽  
Author(s):  
Frank V. Cochran ◽  
Sophia P. Wu ◽  
Wei Wang ◽  
Vikas Nanda ◽  
Jeffery G. Saven ◽  
...  

Science ◽  
1990 ◽  
pp. 973-973
Author(s):  
M. H. Hecht ◽  
J. H. Richardson ◽  
D. C. Richardson ◽  
R. C. Ogden

2014 ◽  
Vol 24 (4) ◽  
pp. 434-445 ◽  
Author(s):  
Grant S. Murphy ◽  
Bharatwaj Sathyamoorthy ◽  
Bryan S. Der ◽  
Mischa C. Machius ◽  
Surya V. Pulavarti ◽  
...  

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