Proteomic determination of the lysine acetylome and phosphoproteome in the rat native inner medullary collecting duct

Kelly A. Hyndman, Chin Rang Yang, Hyun Jun Jung, Ezigbobiara N. Umejiego, Chung Ling Chou, Mark A. Knepper

Research output: Contribution to journalArticle

Abstract

Phosphorylation and lysine (K)-acetylation are dynamic posttranslational modifications of proteins. Previous proteomic studies have identified over 170,000 phosphorylation sites and 15,000 K-acetylation sites in mammals. We recently reported that the inner medullary collecting duct (IMCD), which functions in the regulation of water-reabsorption, via the actions of vasopressin, expresses many of the enzymes that can modulated K-acetylation. The purpose of this study was to determine the K-acetylated or phosphorylated proteins expressed in IMCD cells. Second we questioned whether vasopressin V2 receptor activation significantly affects the IMCD acetylome or phosphoproteome? K-acetylated or serine-, threonine-, or tyrosine-phosphorylated peptides were identified from native rat IMCDs by proteomic analysis with four different enzymes (trypsin, chymotrypsin, ASP-N, or Glu-C) to generate a high-resolution proteome. K-acetylation was identified in 431 unique proteins, and 64% of the K-acetylated sites were novel. The acetylated proteins were expressed in all compartments of the cell and were enriched in pathways including glycolysis and vasopressin-regulated water reabsorption. In the vasopressin-regulated water reabsorption pathway, eight proteins were acetylated, including the novel identification of the basolateral water channel, AQP3, acetylated at K282; 215 proteins were phosphorylated in this IMCD cohort, including AQP2 peptides that were phosphorylated at four serines: 256, 261, 264, and 269. Acute dDAVP did not significantly affect the IMCD acetylome; however, it did significantly affect previously known vasopressin-regulated phosphorylation sites. In conclusion, presence of K-acetylated proteins involved in metabolism, ion, and water transport in the IMCD points to multiple roles of K-acetylation beyond its canonical role in transcriptional regulation.

Original languageEnglish (US)
Pages (from-to)669-679
Number of pages11
JournalPhysiological Genomics
Volume50
Issue number9
DOIs
StatePublished - Sep 2018
Externally publishedYes

Fingerprint

Acetylation
Proteomics
Lysine
Vasopressins
Water
Phosphorylation
Proteins
Serine
Vasopressin Receptors
Aquaporins
Peptides
Ion Transport
Glycolysis
Enzymes
Threonine
Proteome
Post Translational Protein Processing
Tyrosine
Mammals
protein K

Keywords

  • IMCD
  • Lysine acetylation
  • Phosphorylation
  • Vasopressin

ASJC Scopus subject areas

  • Physiology
  • Genetics

Cite this

Proteomic determination of the lysine acetylome and phosphoproteome in the rat native inner medullary collecting duct. / Hyndman, Kelly A.; Yang, Chin Rang; Jung, Hyun Jun; Umejiego, Ezigbobiara N.; Chou, Chung Ling; Knepper, Mark A.

In: Physiological Genomics, Vol. 50, No. 9, 09.2018, p. 669-679.

Research output: Contribution to journalArticle

Hyndman, Kelly A. ; Yang, Chin Rang ; Jung, Hyun Jun ; Umejiego, Ezigbobiara N. ; Chou, Chung Ling ; Knepper, Mark A. / Proteomic determination of the lysine acetylome and phosphoproteome in the rat native inner medullary collecting duct. In: Physiological Genomics. 2018 ; Vol. 50, No. 9. pp. 669-679.
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AU - Hyndman, Kelly A.

AU - Yang, Chin Rang

AU - Jung, Hyun Jun

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AU - Chou, Chung Ling

AU - Knepper, Mark A.

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AB - Phosphorylation and lysine (K)-acetylation are dynamic posttranslational modifications of proteins. Previous proteomic studies have identified over 170,000 phosphorylation sites and 15,000 K-acetylation sites in mammals. We recently reported that the inner medullary collecting duct (IMCD), which functions in the regulation of water-reabsorption, via the actions of vasopressin, expresses many of the enzymes that can modulated K-acetylation. The purpose of this study was to determine the K-acetylated or phosphorylated proteins expressed in IMCD cells. Second we questioned whether vasopressin V2 receptor activation significantly affects the IMCD acetylome or phosphoproteome? K-acetylated or serine-, threonine-, or tyrosine-phosphorylated peptides were identified from native rat IMCDs by proteomic analysis with four different enzymes (trypsin, chymotrypsin, ASP-N, or Glu-C) to generate a high-resolution proteome. K-acetylation was identified in 431 unique proteins, and 64% of the K-acetylated sites were novel. The acetylated proteins were expressed in all compartments of the cell and were enriched in pathways including glycolysis and vasopressin-regulated water reabsorption. In the vasopressin-regulated water reabsorption pathway, eight proteins were acetylated, including the novel identification of the basolateral water channel, AQP3, acetylated at K282; 215 proteins were phosphorylated in this IMCD cohort, including AQP2 peptides that were phosphorylated at four serines: 256, 261, 264, and 269. Acute dDAVP did not significantly affect the IMCD acetylome; however, it did significantly affect previously known vasopressin-regulated phosphorylation sites. In conclusion, presence of K-acetylated proteins involved in metabolism, ion, and water transport in the IMCD points to multiple roles of K-acetylation beyond its canonical role in transcriptional regulation.

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