Role of aspartic acid residues D87 and D89 in APS kinase domain of human 3 '-phosphoadenosine 5 '-phosphosulfate synthase 1 and 2b: A commonality with phosphatases/kinases

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11320%2F21%3A10438876" target="_blank" >RIV/00216208:11320/21:10438876 - isvavai.cz</a>

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=h6T4udGaIG" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=h6T4udGaIG</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bbrep.2021.101155" target="_blank" >10.1016/j.bbrep.2021.101155</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Role of aspartic acid residues D87 and D89 in APS kinase domain of human 3 '-phosphoadenosine 5 '-phosphosulfate synthase 1 and 2b: A commonality with phosphatases/kinases

Popis výsledku v původním jazyce

3&apos;-phosphoadenosine 5&apos;-phosphosulfate (PAPS) is synthesized in two steps by PAPS synthase (PAPSS). PAPSS is comprised of ATP sulfurylase (ATPS) and APS kinase (APSK) domain activities. ATPS combines inorganic sulfate with a-phosphoryl of ATP to form adenosine 5&apos;-phosphosulfate (APS) and PPi. In the second step APS is phosphorylated at 3&apos;-OH using another mole of ATP to form PAPS and ADP catalyzed by APSK. The transfer of gamma-phosphoryl from ATP onto 3&apos;-OH requires Mg-2(+) and purported to involve residues D(87)GD(89)N. We report that mutation of either aspartic residue to alanine completely abolishes APSK activity in PAPS formation. PAPSS is an, unique enzyme that binds to four different nucleotides: ATP and APS on both ATPS and APSK domains and ADP and PAPS exclusively on the APSK domain. The thermodynamic binding and the catalytic interplay must be very tightly controlled to form the end-product PAPS in the forward direction. Though APS binds to ATPS and APSK, in ATPS domain, the APS is a product and for APSK it is a substrate. DGDN motif is absent in ATPS and present in APSK. Mutation of D-87 and D-89 did not hamper ATPS activity however abolished APSK activity severely. Thus, D(87)GD(89)N region is required for stabilization of Mg2+-ATP, in the process of splitting the 7-phosphoryl from ATP and transfer of 7-phosphoryl onto 3&apos;-OH of APS to form PAPS a process that cannot be achieved by ATPS domain. In addition, gamma P-32-ATP, trapped phosphoryl enzyme intermediate more with PAPSS2 than with PAPSS1. This suggests inherent active site residues could control novel catalytic differences. Molecular docking studies of hPAPSS1 with ATP + Mg2+ and APS of wild type and mutants supports the experimental results.

Název v anglickém jazyce

Role of aspartic acid residues D87 and D89 in APS kinase domain of human 3 '-phosphoadenosine 5 '-phosphosulfate synthase 1 and 2b: A commonality with phosphatases/kinases

Popis výsledku anglicky

3&apos;-phosphoadenosine 5&apos;-phosphosulfate (PAPS) is synthesized in two steps by PAPS synthase (PAPSS). PAPSS is comprised of ATP sulfurylase (ATPS) and APS kinase (APSK) domain activities. ATPS combines inorganic sulfate with a-phosphoryl of ATP to form adenosine 5&apos;-phosphosulfate (APS) and PPi. In the second step APS is phosphorylated at 3&apos;-OH using another mole of ATP to form PAPS and ADP catalyzed by APSK. The transfer of gamma-phosphoryl from ATP onto 3&apos;-OH requires Mg-2(+) and purported to involve residues D(87)GD(89)N. We report that mutation of either aspartic residue to alanine completely abolishes APSK activity in PAPS formation. PAPSS is an, unique enzyme that binds to four different nucleotides: ATP and APS on both ATPS and APSK domains and ADP and PAPS exclusively on the APSK domain. The thermodynamic binding and the catalytic interplay must be very tightly controlled to form the end-product PAPS in the forward direction. Though APS binds to ATPS and APSK, in ATPS domain, the APS is a product and for APSK it is a substrate. DGDN motif is absent in ATPS and present in APSK. Mutation of D-87 and D-89 did not hamper ATPS activity however abolished APSK activity severely. Thus, D(87)GD(89)N region is required for stabilization of Mg2+-ATP, in the process of splitting the 7-phosphoryl from ATP and transfer of 7-phosphoryl onto 3&apos;-OH of APS to form PAPS a process that cannot be achieved by ATPS domain. In addition, gamma P-32-ATP, trapped phosphoryl enzyme intermediate more with PAPSS2 than with PAPSS1. This suggests inherent active site residues could control novel catalytic differences. Molecular docking studies of hPAPSS1 with ATP + Mg2+ and APS of wild type and mutants supports the experimental results.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10301 - Atomic, molecular and chemical physics (physics of atoms and molecules including collision, interaction with radiation, magnetic resonances, Mössbauer effect)

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Biochemistry and Biophysics Reports [online]

ISSN

2405-5808

e-ISSN

—

Svazek periodika

28

Číslo periodika v rámci svazku

5

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

9

Strana od-do

101155

Kód UT WoS článku

000732710600008

EID výsledku v databázi Scopus

2-s2.0-85122798048