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Global protein dephosphorylation patterns alter the performances of microbial cell factories

ABSTRACT: Global regulation of protein phosphorylation and dephosphorylation plays crucial roles in various cellular processes, serving as a fundamental mechanism for regulating protein function, cell metabolic performance and stress response. In response to differences in nutrition, alterations in the metabolic flux are accompanied by the changes in the phosphorylation levels of enzymes. However, the stress-driven large-scale protein phosphorylation in cells and the underlying principles remain unclear. Here, we report a global switchable phosphorylation pattern of yeast cells in response to environmental variations, which is governed reversibly with one bifunction phosphatase regulator, Ykr075c. We found this regulator could change the phosphorylation pattern of proteins, covering carbon metabolism and signal transduction, and thus greatly enhancing the growth and cell metabolism under the heat stress. As reasons, it dephosphorylates glycogen synthesis enzymes and affects enzyme activities as well, and it also mediates the dephosphorylation of several protein kinases and glucose metabolism-related transcription factors, thereby elevating carbon metabolism at the transcriptional level. Based on these findings, we significantly enhanced the growth and production of yeast cells, exhibiting the universality across variable products and diverse species and offering novel insights into the regulation of protein phosphorylation for designing microbial cell factories.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast) Saccharomyces Cerevisiae

SUBMITTER: Lei Qin

LAB HEAD: Chun Li

PROVIDER: PXD056937 | Pride | 2024-10-23

REPOSITORIES: Pride

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			Action	DRS
	F1_yeast_phospho_BY4742_38_1.raw	Raw
	F2_yeast_phospho_BY4742_38_2.raw	Raw
	F3_yeast_phospho_BY4742_30_1.raw	Raw
	F4_yeast_phospho_BY4742_30_2.raw	Raw
	F5_yeast_phospho_YKR075C_38_1.raw	Raw

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Project description:This study aims to analyze and compare the post-translational modification differences between human plasma- and genetic recombination-derived human serum albumin (HSA) in clinical treatments in China. 6 different post-translational modifications, including acetylation, succinylation, crotonylation, phosphorylation, beta-hydroxybutyrylation, and lactylation, were detected using pan-specific antibodies through Western blot. The samples, consisting of human plasma-derived HSA (pHSA) from six different manufacturers and recombinant HSA (rHSA) expressed in yeast and oryza sativa, underwent detection of types of post-translational modifications. 4D label-free PTMs quantitative proteomic analysis was conducted to detect N-glycosylation and the above-mentioned post-translational modifications in pHSA and rHSA samples, and analyze the differences in modification sites and levels. Through Western blot analysis, all six pHSA and two rHSA samples were positive in the band of albumin (66.5 kDa) for the six post-translational modifications. Further analysis using 4D label-free PTMs quantitative proteomics revealed 25 (29) acetylated, 30 (32) succinylated, 41 (50) malonylated, 15 (23) phosphorylated, 36 (30) beta-hydroxybutyrylated, and 27 (34) lactylated modification sites in pHSA and rHSA samples and no N-glycosylation modification sites. The analysis results identified 1 acetylation (ALB_K160), 2 beta-hydroxybutyrylation (ALB_K569, ALB_K426), 3 crotonylation (ALB_K264, ALB_K581, ALB_K560), and 15 phosphorylation specific modification sites in pHSA, as well as 3 crotonylation (ALB_K560, ALB_K562, ALB_K75), 1 succinylation (ALB_K490), and 23 phosphorylation specific modification sites in rHSA. In pHSA (rHSA), 2 (6) acetylation, 10 (12) succinylation, 0 (9) crotonylation, 1 (9) phosphorylation, 6 (0) beta-hydroxybutyrylation, and 0 (7) lactylation specific modification sites were found. Additionally, in the common modification sites of pHSA and rHSA, pHSA showed up-regulation of amberylation (16:1) and beta-hydroxybutyrylation (12:2) in more sites, and up-regulation of acetylation (7:11), crotonylation (2:11), phosphorylation (1:8), and lactylation (1:14) in less sites compared to rHSA. In clinical treatment, the used pHSA and rHSA in China generally exhibit acetylation, succinylation, crotonylation, phosphorylation, beta-hydroxybutyrylation, and lactylation, and there are differences in the site characteristics and modification levels of these modifications between pHSA and rHSA. Further experimental investigations are needed to explore the impact of post-translational modification differences on the biological function, efficacy, and safety of pHSA and rHSA.

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Global protein dephosphorylation patterns alter the performances of microbial cell factories

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