Enzyme activity; Phosphofructokinase; Protein acetylation; Protein phosphorylation; Food Science; Biochemistry
Abstract :
[en] Phosphofructokinase (PFK) is a key glycolytic rate-limiting enzyme that affects final meat quality through glycolysis regulation. PFK occurs protein phosphorylation and acetylation in postmortem meat, but how they affect PFK activity has not been well studied. Phosphorylation and acetylation of PFK were adjusted in an in vitro system by adding protein kinase A (PKA) and P300/CBP-associated factor (PCAF), respectively. PFK activity significantly decreased after increasing its phosphorylation level by PKA (P < 0.05). PCAF increased PFK acetylation level, but did not significantly change its activity (P > 0.05). Five PFK variants including wtPFK (wild type), PFK_T704D (T704 phosphorylation), PFK_T704A (T704 dephosphorylation), PFK_K678Q (K678 acetylation) and PFK_K678R (K678 acetylation) were constructed to investigated the effect of T704 and K678 on enzyme activity. Kinetic analysis showed a higher Km, lower Vmax, and lower catalytic efficiency of PFK_T704D on fructose 6-phosphate (F6P), suggesting that T704 phosphorylation decreased PFK activity, possibly by weakening the affinity between F6P and PFK. The changes of PFK variants on ATP were slightly lower than those of PFK variants on F6P. The present study identifies T704 as a possibly vital site for PFK activity regulation and provides theoretical support for the development of meat quality preservation technology.
Disciplines :
Agriculture & agronomy
Author, co-author :
Ren, Chi ; Université de Liège - ULiège > TERRA Research Centre ; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Zhao, Xinran; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Schroyen, Martine ; Université de Liège - ULiège > TERRA Research Centre > Animal Sciences (AS)
Li, Xin ; Université de Liège - ULiège > TERRA Research Centre ; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Hou, Chengli; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Wang, Zhenyu; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Zhang, Dequan; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agro-products Quality & Safety in Harvest, Storage, Transportation, Management and Control, Ministry of Agriculture and Rural Affairs, Beijing, China
Language :
English
Title :
The effect of protein phosphorylation and acetylation on phosphofructokinase in lamb
NSCF - National Natural Science Foundation of China
Funding text :
This study was financially supported by the National Natural Science Foundation of China (32030086, 32072245), Agricultural Science and Technology Innovation Program (CAAS-ASTIP-2023-IFST-03) in China.
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