Effects of Phosphorylation on Titin Degradation at Different Ca2+ Concentrations Incubation in vitro

Abstract

Abstract: This study aimed to investigate effects of phosphorylation on titin degradation at different Ca2+ concentrations incubation in vitro. Myofibrillar proteins were extracted from ovine right longissimus lumborum muscles 30 min after the slaughter. Low molecular weight proteins (<150 kDa) was removed through size exclusion chromatography to obtain the crude titin. To verify the presence of titin, the images of the crude titin was explored with atomic force microscope (AFM). Protein kinase A (PKA) and alkaline phosphatase (AP) were added to titin in vitro to alter its phosphorylation level for 30 min at 30 ℃, while the protein dissolution buffer was added to the control group. Samples were then incubated with μ-calpain at 4 ℃ for 2 days at the Ca2+ concentration of 0.05 mM and 2 mM, respectively. The results showed that pH values of the incubation system from three groups had no significant difference (P>0.05). The relative phosphorylation level of titin from the PKA group was higher than that from the control group, which had a higher relative phosphorylation level than in the AP group (P<0.05). The degradation rate of titin was faster at the Ca2+ concentration of 2 mM, compared with that at the Ca2+ concentration of 0.05 mM. Titin in the AP group degraded earlier than that in the PKA and control group at the same Ca2+ concentration. These observations demonstrated that the phosphorylation of titin in vitro was significantly increased by PKA, while decreased by AP. With the increase of calcium concentration, the degradation rate of titin became faster. The dephosphorylation of titin promoted the titin degradation.

Key words: Titin, Protein kinase A, Alkaline phosphatase, Phosphorylation, Degradation

CLC Number:

TS251.1

Xin Li Zhang Sheqi. Effects of Phosphorylation on Titin Degradation at Different Ca2+ Concentrations Incubation in vitro[J]. FOOD SCIENCE, 0, (): 0-0.

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