Enzyme catalysis prior to aromatic residues: Reverse engineering of a dephospho‐CoA kinase
Identifikátory výsledku
Kód výsledku v IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F21%3A00541647" target="_blank" >RIV/61388963:_____/21:00541647 - isvavai.cz</a>
Nalezeny alternativní kódy
RIV/00216208:11310/21:10439001 RIV/00216224:14740/21:00124510
Výsledek na webu
<a href="https://doi.org/10.1002/pro.4068" target="_blank" >https://doi.org/10.1002/pro.4068</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/pro.4068" target="_blank" >10.1002/pro.4068</a>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
Enzyme catalysis prior to aromatic residues: Reverse engineering of a dephospho‐CoA kinase
Popis výsledku v původním jazyce
The wide variety of protein structures and functions results from the diverse properties of the 20 canonical amino acids. The generally accepted hypothesis is that early protein evolution was associated with enrichment of a primordial alphabet, thereby enabling increased protein catalytic efficiencies and functional diversification. Aromatic amino acids were likely among the last additions to genetic code. The main objective of this study was to test whether enzyme catalysis can occur without the aromatic residues (aromatics) by studying the structure and function of dephospho‐CoA kinase (DPCK) following aromatic residue depletion. We designed two variants of a putative DPCK from Aquifex aeolicus by substituting (a) Tyr, Phe and Trp or (b) all aromatics (including His). Their structural characterization indicates that substituting the aromatics does not markedly alter their secondary structures but does significantly loosen their side chain packing and increase their sizes. Both variants still possess ATPase activity, although with 150–300 times lower efficiency in comparison with the wild‐type phosphotransferase activity. The transfer of the phosphate group to the dephospho‐CoA substrate becomes heavily uncoupled and only the His‐containing variant is still able to perform the phosphotransferase reaction. These data support the hypothesis that proteins in the early stages of life could support catalytic activities, albeit with low efficiencies. An observed significant contraction upon ligand binding is likely important for appropriate organization of the active site. Formation of firm hydrophobic cores, which enable the assembly of stably structured active sites, is suggested to provide a selective advantage for adding the aromatic residues.
Název v anglickém jazyce
Enzyme catalysis prior to aromatic residues: Reverse engineering of a dephospho‐CoA kinase
Popis výsledku anglicky
The wide variety of protein structures and functions results from the diverse properties of the 20 canonical amino acids. The generally accepted hypothesis is that early protein evolution was associated with enrichment of a primordial alphabet, thereby enabling increased protein catalytic efficiencies and functional diversification. Aromatic amino acids were likely among the last additions to genetic code. The main objective of this study was to test whether enzyme catalysis can occur without the aromatic residues (aromatics) by studying the structure and function of dephospho‐CoA kinase (DPCK) following aromatic residue depletion. We designed two variants of a putative DPCK from Aquifex aeolicus by substituting (a) Tyr, Phe and Trp or (b) all aromatics (including His). Their structural characterization indicates that substituting the aromatics does not markedly alter their secondary structures but does significantly loosen their side chain packing and increase their sizes. Both variants still possess ATPase activity, although with 150–300 times lower efficiency in comparison with the wild‐type phosphotransferase activity. The transfer of the phosphate group to the dephospho‐CoA substrate becomes heavily uncoupled and only the His‐containing variant is still able to perform the phosphotransferase reaction. These data support the hypothesis that proteins in the early stages of life could support catalytic activities, albeit with low efficiencies. An observed significant contraction upon ligand binding is likely important for appropriate organization of the active site. Formation of firm hydrophobic cores, which enable the assembly of stably structured active sites, is suggested to provide a selective advantage for adding the aromatic residues.
Klasifikace
Druh
J<sub>imp</sub> - Článek v periodiku v databázi Web of Science
CEP obor
—
OECD FORD obor
10608 - Biochemistry and molecular biology
Návaznosti výsledku
Projekt
Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.
Návaznosti
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Ostatní
Rok uplatnění
2021
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
Protein Science
ISSN
0961-8368
e-ISSN
1469-896X
Svazek periodika
30
Číslo periodika v rámci svazku
5
Stát vydavatele periodika
US - Spojené státy americké
Počet stran výsledku
13
Strana od-do
1022-1034
Kód UT WoS článku
000632928300001
EID výsledku v databázi Scopus
2-s2.0-85103210737