Myocellular creatine and creatine transporter serine phosphorylation after starvation

Chun Rui Zhao, Lihong Shang, Weiyang Wang, Danny O. Jacobs

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Background. Myocellular creatine, which is critically important for normal energy metabolism, increases in rat gastrocnemius muscle after starvation via unknown mechanisms. Creatine (Cr) uptake across plasma membranes is governed by a single, specific transporter (CrTr) that shares 50% amino acid sequence identity with GABA/choline/betaine transporters whose functions are modulated by phosphorylation. Methods. Gastrocnemius muscle was collected from adult male Sprague-Dawley (225-250 g) rats that were randomized to receive normal rat chow and distilled water ad libitum (CTL) or distilled water alone for 4 days (STV). Total Cr, phosphocreatine (PCr), free Cr, and ATP were measured luminometrically. CrTr protein expression and protein serine and tyrosine phosphorylation and mRNA expression were determined using immunoprecipitation and quantitative Western blotting and reverse transcription polymerase chain reaction (RT-PCR) analyses, respectively. Guanidinoacetate methyltransferase (GAMT) activity, guanidinoacetic acid (GAA) content, creatine kinase (CK) activity, and creatinine (Crn) content were assayed luminometrically or spectrophotometrically. Creatine transporter uptake activity was also measured in skeletal muscle membrane vesicles. Data were analyzed by t test. Results. Total Cr and free Cr increased 26 and 280% in STV (32.3 ± 1.0 and 12.9 ± 1.4 vs 25.7 ± 1.1 and 3.4 ± 0.9 μmol/g wet wt, mean ± SEM, respectively, P < 0.01) whereas PCr content decreased 18% (18.6 ± 0.8 vs 22.8 ± 0.9 μmol/g wet wt, STV vs CTL P < 0.05). CrTr protein and mRNA expression, ATP, GAA, CK, GAMT, and protein tyrosine phosphorylation of CrTr were not significantly different between the two groups. However, protein serine phosphorylation of CrTr was significantly reduced by 30% (P < 0.05) and creatine uptake activity was significantly increased (P < 0.05) in starved animals. Conclusion. Increases in myocellular creatine content after starvation are associated with reduced serine phosphorylation of the creatine transporter.

Original languageEnglish (US)
Pages (from-to)10-16
Number of pages7
JournalJournal of Surgical Research
Volume105
Issue number1
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Creatine
Starvation
Serine
Phosphorylation
Guanidinoacetate N-Methyltransferase
Skeletal Muscle
Phosphocreatine
Creatine Kinase
Proteins
Tyrosine
Adenosine Triphosphate
Messenger RNA
creatine transporter
Water
Immunoprecipitation
gamma-Aminobutyric Acid
Energy Metabolism
Reverse Transcription
Amino Acid Sequence
Creatinine

Keywords

  • Creatine kinase
  • Creatine transporter
  • Creatine uptake
  • Guanidinoacetate methyltransferase
  • Guanidinoacetic acid
  • Luminometry
  • Protein phosphorylation
  • Skeletal muscle membrane vesicle
  • Starvation

ASJC Scopus subject areas

  • Surgery

Cite this

Myocellular creatine and creatine transporter serine phosphorylation after starvation. / Zhao, Chun Rui; Shang, Lihong; Wang, Weiyang; Jacobs, Danny O.

In: Journal of Surgical Research, Vol. 105, No. 1, 2002, p. 10-16.

Research output: Contribution to journalArticle

Zhao, Chun Rui ; Shang, Lihong ; Wang, Weiyang ; Jacobs, Danny O. / Myocellular creatine and creatine transporter serine phosphorylation after starvation. In: Journal of Surgical Research. 2002 ; Vol. 105, No. 1. pp. 10-16.
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title = "Myocellular creatine and creatine transporter serine phosphorylation after starvation",
abstract = "Background. Myocellular creatine, which is critically important for normal energy metabolism, increases in rat gastrocnemius muscle after starvation via unknown mechanisms. Creatine (Cr) uptake across plasma membranes is governed by a single, specific transporter (CrTr) that shares 50{\%} amino acid sequence identity with GABA/choline/betaine transporters whose functions are modulated by phosphorylation. Methods. Gastrocnemius muscle was collected from adult male Sprague-Dawley (225-250 g) rats that were randomized to receive normal rat chow and distilled water ad libitum (CTL) or distilled water alone for 4 days (STV). Total Cr, phosphocreatine (PCr), free Cr, and ATP were measured luminometrically. CrTr protein expression and protein serine and tyrosine phosphorylation and mRNA expression were determined using immunoprecipitation and quantitative Western blotting and reverse transcription polymerase chain reaction (RT-PCR) analyses, respectively. Guanidinoacetate methyltransferase (GAMT) activity, guanidinoacetic acid (GAA) content, creatine kinase (CK) activity, and creatinine (Crn) content were assayed luminometrically or spectrophotometrically. Creatine transporter uptake activity was also measured in skeletal muscle membrane vesicles. Data were analyzed by t test. Results. Total Cr and free Cr increased 26 and 280{\%} in STV (32.3 ± 1.0 and 12.9 ± 1.4 vs 25.7 ± 1.1 and 3.4 ± 0.9 μmol/g wet wt, mean ± SEM, respectively, P < 0.01) whereas PCr content decreased 18{\%} (18.6 ± 0.8 vs 22.8 ± 0.9 μmol/g wet wt, STV vs CTL P < 0.05). CrTr protein and mRNA expression, ATP, GAA, CK, GAMT, and protein tyrosine phosphorylation of CrTr were not significantly different between the two groups. However, protein serine phosphorylation of CrTr was significantly reduced by 30{\%} (P < 0.05) and creatine uptake activity was significantly increased (P < 0.05) in starved animals. Conclusion. Increases in myocellular creatine content after starvation are associated with reduced serine phosphorylation of the creatine transporter.",
keywords = "Creatine kinase, Creatine transporter, Creatine uptake, Guanidinoacetate methyltransferase, Guanidinoacetic acid, Luminometry, Protein phosphorylation, Skeletal muscle membrane vesicle, Starvation",
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T1 - Myocellular creatine and creatine transporter serine phosphorylation after starvation

AU - Zhao, Chun Rui

AU - Shang, Lihong

AU - Wang, Weiyang

AU - Jacobs, Danny O.

PY - 2002

Y1 - 2002

N2 - Background. Myocellular creatine, which is critically important for normal energy metabolism, increases in rat gastrocnemius muscle after starvation via unknown mechanisms. Creatine (Cr) uptake across plasma membranes is governed by a single, specific transporter (CrTr) that shares 50% amino acid sequence identity with GABA/choline/betaine transporters whose functions are modulated by phosphorylation. Methods. Gastrocnemius muscle was collected from adult male Sprague-Dawley (225-250 g) rats that were randomized to receive normal rat chow and distilled water ad libitum (CTL) or distilled water alone for 4 days (STV). Total Cr, phosphocreatine (PCr), free Cr, and ATP were measured luminometrically. CrTr protein expression and protein serine and tyrosine phosphorylation and mRNA expression were determined using immunoprecipitation and quantitative Western blotting and reverse transcription polymerase chain reaction (RT-PCR) analyses, respectively. Guanidinoacetate methyltransferase (GAMT) activity, guanidinoacetic acid (GAA) content, creatine kinase (CK) activity, and creatinine (Crn) content were assayed luminometrically or spectrophotometrically. Creatine transporter uptake activity was also measured in skeletal muscle membrane vesicles. Data were analyzed by t test. Results. Total Cr and free Cr increased 26 and 280% in STV (32.3 ± 1.0 and 12.9 ± 1.4 vs 25.7 ± 1.1 and 3.4 ± 0.9 μmol/g wet wt, mean ± SEM, respectively, P < 0.01) whereas PCr content decreased 18% (18.6 ± 0.8 vs 22.8 ± 0.9 μmol/g wet wt, STV vs CTL P < 0.05). CrTr protein and mRNA expression, ATP, GAA, CK, GAMT, and protein tyrosine phosphorylation of CrTr were not significantly different between the two groups. However, protein serine phosphorylation of CrTr was significantly reduced by 30% (P < 0.05) and creatine uptake activity was significantly increased (P < 0.05) in starved animals. Conclusion. Increases in myocellular creatine content after starvation are associated with reduced serine phosphorylation of the creatine transporter.

AB - Background. Myocellular creatine, which is critically important for normal energy metabolism, increases in rat gastrocnemius muscle after starvation via unknown mechanisms. Creatine (Cr) uptake across plasma membranes is governed by a single, specific transporter (CrTr) that shares 50% amino acid sequence identity with GABA/choline/betaine transporters whose functions are modulated by phosphorylation. Methods. Gastrocnemius muscle was collected from adult male Sprague-Dawley (225-250 g) rats that were randomized to receive normal rat chow and distilled water ad libitum (CTL) or distilled water alone for 4 days (STV). Total Cr, phosphocreatine (PCr), free Cr, and ATP were measured luminometrically. CrTr protein expression and protein serine and tyrosine phosphorylation and mRNA expression were determined using immunoprecipitation and quantitative Western blotting and reverse transcription polymerase chain reaction (RT-PCR) analyses, respectively. Guanidinoacetate methyltransferase (GAMT) activity, guanidinoacetic acid (GAA) content, creatine kinase (CK) activity, and creatinine (Crn) content were assayed luminometrically or spectrophotometrically. Creatine transporter uptake activity was also measured in skeletal muscle membrane vesicles. Data were analyzed by t test. Results. Total Cr and free Cr increased 26 and 280% in STV (32.3 ± 1.0 and 12.9 ± 1.4 vs 25.7 ± 1.1 and 3.4 ± 0.9 μmol/g wet wt, mean ± SEM, respectively, P < 0.01) whereas PCr content decreased 18% (18.6 ± 0.8 vs 22.8 ± 0.9 μmol/g wet wt, STV vs CTL P < 0.05). CrTr protein and mRNA expression, ATP, GAA, CK, GAMT, and protein tyrosine phosphorylation of CrTr were not significantly different between the two groups. However, protein serine phosphorylation of CrTr was significantly reduced by 30% (P < 0.05) and creatine uptake activity was significantly increased (P < 0.05) in starved animals. Conclusion. Increases in myocellular creatine content after starvation are associated with reduced serine phosphorylation of the creatine transporter.

KW - Creatine kinase

KW - Creatine transporter

KW - Creatine uptake

KW - Guanidinoacetate methyltransferase

KW - Guanidinoacetic acid

KW - Luminometry

KW - Protein phosphorylation

KW - Skeletal muscle membrane vesicle

KW - Starvation

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