Role of PEPT2 in glycylsarcosine transport in astrocyte and glioma cultures

Xiang, Jianming; Chiang, Pei Pei; Hu, Yongjun; Smith, David Eugene; Keep, Richard F.

doi:10.1016/j.neulet.2005.11.037

Cited by 15 publications

(19 citation statements)

References 19 publications

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“…In the cultured kidney cell line SKPT, and in rat renal BBMV, the K m for Gly-sar uptake is around 50 μM [3,8,19,21]. In cultured rat neonatal astrocytes, the K m for Gly-sar uptake was 222 μM [27]. The small discrepancies between these values for Glysar uptake by PEPT2 in several preparations including that measured in the current study (Fig.…”

Section: Discussionmentioning

confidence: 57%

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Demonstration of functional dipeptide transport with expression of PEPT2 in guinea pig cardiomyocytes

Lin

King

2006

Pflugers Arch - Eur J Physiol

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The transporters PEPT1 and PEPT2 accept a broad spectrum of substrates including small, naturally occurring peptides and peptidomimetic drugs. This study aimed to investigate for the first time whether these transporters are expressed and active in isolated cardiomyocytes. PEPT1/PEPT2 expression in rat kidney (positive control), guinea pig kidney and cardiomyocytes were investigated by reverse transcription polymerase chain reaction. L-Glycyl-L-[(14)C]sarcosine (Gly-sar) uptake was characterised using freshly isolated suspensions of adult male guinea pig cardiomyocytes. PEPT2-specific primers recognised mRNA of appropriate size and sequence in cardiomyocytes and kidney, whilst PEPT1 was expressed in the kidney only. The initial uptake (30 s) of 200 microM Gly-sar was dependent on extracellular pH with a maximum at pH 6.0 (237.8 +/- 12.2 pmol/microl) and a minimum at pH 8.0 (72.1 +/- 13.4 pmol/microl, n = 6 +/- SE, p < 0.01, T test). The K (m) and V (max) of Gly-sar uptake at pH 6.0 were 495.5 +/- 69.6 microM and 1470.5 +/- 69.6 pmol microl(-1) min(-1). The addition of 10 mM fosinopril, cefadroxil, carnosine, cyclacillin or a variety of L-amino acid containing dipeptides/tripeptides significantly reduced Gly-sar uptake. Gly-sar uptake was not affected by 10 mM D-ala-D-ala, glycine or sarcosine. These results support the presence of a functional dipeptide transporter in isolated cardiomyocytes, with accompanying expression of PEPT2.

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Section: Discussionmentioning

confidence: 57%

“…[3,7,8,17,23,27]), although not in heart. The consensus amongst these studies is that, regardless of the transporter isoform involved, Gly-sar uptake is strongly dependent on an inward proton gradient.…”

Section: Discussionmentioning

confidence: 97%

Demonstration of functional dipeptide transport with expression of PEPT2 in guinea pig cardiomyocytes

Lin

King

2006

Pflugers Arch - Eur J Physiol

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“…As stated earlier, FBS is a complex mixture of nutrients conventionally used for culturing eukaryotic cells. It has also been used as a trypsin inhibitor in some previous studies (Xiang et al 2006;Wang et al 2008bWang et al , 2009) because it contains a-2-macroglobulin, a large plasma protein (60 to 80kDa) (Dangott and Cunningham 1982) capable of inactivating different types of proteinases (Barrett and Starkey 1973). In addition, another protein in FBS, a-1 antitrypsin, which has a mass of 52 kDa, was reportedly able to specifically and irreversibly inactivate the enzyme trypsin in vitro (Hercz 1986;Ozer 1998).…”

Section: Discussion and Summarymentioning

confidence: 99%

“…In previous studies, the trypsin activity has also been inhibited by steeping the trypsin-treated specimens in a saline buffer with ethylenediamine tetraacetic acid (EDTA) and different protease inhibitors, including benzamidine hydrochloride, N-ethylmaleimide (NEM) and phenylmethylsulfonyl fluoride (PMSF) Rieppo et al 2003) or fetal bovine serum (FBS) (Xiang et al 2006). Fetal bovine serum is a very complex mixture of nutrients including proteins, growth factors, vitamins, lipids, salts, minerals and amino acids, allowing for culturing of eukaryotic cells.…”

Section: Introductionmentioning

confidence: 99%

Real-Time Electro-Mechano-Acoustic Imaging for Monitoring Interactions Between Trypsin and Different Inhibitors in Articular Cartilage

Zheng

Wang

Butt

2011

Ultrasound in Medicine & Biology

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“…It is widely expressed in brain, kidney, lung, eye and mammary gland [2,3]. The functional activity of PEPT2 has been studied using a variety of substrates including the synthetic dipeptide glycylsarcosine (GlySar) [4–6], endogenous peptidomimetics (e.g., 5-aminolevulinic acid) [7,8], neuropeptides (e.g., carnosine and kyotorphin) [9–11], as well as peptide-like drugs (e.g., cefadroxil) [12,13]. In kidney, PEPT2 is expressed at the apical membrane of proximal tubule epithelial cells where it plays an important role in the reabsorption of substrates from urine, thereby limiting renal clearance [14].…”

Section: Introductionmentioning

confidence: 99%

Influence of peptide transporter 2 (PEPT2) on the distribution of cefadroxil in mouse brain: A microdialysis study

Chen

Keep

Liang

et al. 2017

Biochemical Pharmacology

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Peptide transporter 2 (PEPT2) is a high-affinity low-capacity transporter belonging to the proton-coupled oligopeptide transporter family. Although many aspects of PEPT2 structure-function are known, including its localization in choroid plexus and neurons, its regional activity in brain, especially extracellular fluid (ECF), is uncertain. In this study, the pharmacokinetics and regional brain distribution of cefadroxil, a β-lactam antibiotic and PEPT2 substrate, were investigated in wildtype and Pept2 null mice using in vivo intracerebral microdialysis. Cefadroxil was infused intravenously over 4 hours at 0.15 mg/min/kg, and samples obtained from plasma, brain ECF, cerebrospinal fluid (CSF) and brain tissue. A permeability-surface area experiment was also performed in which 0.15 mg/min/kg cefadroxil was infused intravenously for 10 min, and samples obtained from plasma and brain tissues. Our results showed that PEPT2 ablation significantly increased the brain ECF and CSF levels of cefadroxil (2- to 2.5-fold). In contrast, there were no significant differences between wildtype and Pept2 null mice in the amount of cefadroxil in brain cells. The unbound volume of distribution of cefadroxil in brain was 60% lower in Pept2 null mice indicating an uptake function for PEPT2 in brain cells. Finally, PEPT2 did not affect the influx clearance of cefadroxil, thereby, ruling out differences between the two genotypes in drug entry across the blood-brain barriers. These findings demonstrate, for the first time, the impact of PEPT2 on brain ECF as well as the known role of PEPT2 in removing peptide-like drugs, such as cefadroxil, from the CSF to blood.

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Role of PEPT2 in glycylsarcosine transport in astrocyte and glioma cultures

Cited by 15 publications

References 19 publications

Demonstration of functional dipeptide transport with expression of PEPT2 in guinea pig cardiomyocytes

Demonstration of functional dipeptide transport with expression of PEPT2 in guinea pig cardiomyocytes

Real-Time Electro-Mechano-Acoustic Imaging for Monitoring Interactions Between Trypsin and Different Inhibitors in Articular Cartilage

Influence of peptide transporter 2 (PEPT2) on the distribution of cefadroxil in mouse brain: A microdialysis study

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