Uphill transport of beta-alanine in intestinal brush-border membrane vesicles

Miyamoto, Y.; Nakamura, H.; Hoshi, Takeshi; Ganapathy, V.; Leibach, F. H.

doi:10.1152/ajpgi.1990.259.3.g372

Cited by 22 publications

(18 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The kinetical data of the p-alanine transport by muscle cells also compare well with the data reported for other systems such as the high-affinity low-capacity p-alanine uptake system of renal [4] and intestinal [7] brush border membranes.…”

Section: Discussionsupporting

confidence: 82%

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

Bakardjiev¹,

Bauer²

1994

European Journal of Biochemistry

View full text Add to dashboard Cite

Uptake of p-alanine and synthesis of carnosine (P-alanyl-histidine) could be demonstrated in primary cell cultures derived from embryonic chick pectoral muscle. Concomitant with the morphological changes, cessation of cell division and the induction of creatine kinase, a rapid increase in the rate of p-alanine uptake and also in the rate of carnosine synthesis could be observed. The uptake of p-alanine is sodium and chloride dependent and obeys Michaelis-Menten kinetics with K, values of about 40 pM that are essentially identical for myoblasts and myotubes. In contrast, V,, increases considerably during differentiation. The p-alanine transport system is highly specific for p-amino acids and exhibits a substantial anion dependency (Cl-> J-> CSN-> SO:-). Stoichiometric studies suggest that the transport of one p-alanine molecule involves two sodium ions and one chloride ion. This ratio is not altered by the process of cell differentiation.Carnosine (P-alanyl-histidine) and structurally related peptides (w-aminoacyl amino acids) are well-known constituents of excitable tissues [l, 21. In some muscles of mammals and birds, these peptides even represent the major nonproteinaceous constituents. Since p-alanine is predominantly synthesized by the liver as a final metabolite of the pyrimidine bases uracil and thymine but not by muscle cells directly [3], muscle cells must be equipped with a highly efficient transport system for this amino acid. So far, however, uptake of p-alanine from the peripheral circulation has been extensively studied in kidney [4], brain [5] and, to some extent, also in other tissues [6, 71, but surprisingly not yet in skeletal muscle. Here, we report on the uptake of p-alanine and the synthesis of carnosine by primary cultures of chick pectoral muscle. MATERIALS AND METHODSHam's F12 nutrient mixture and horse serum were obtained from Gibco. Cells were grown in cell-culture dishes (Nunc) or flasks (Falcon). 120 Ci/mmol p- Transferrin, bovine serum albumin, phosphoglycerate kinase, hexokinase, glucose-6-phosphate dehydrogenase, creatine phosphate, ADP", NADP", P',P5-diadenosine-5'-pentaphosphate, N-acetyl-L-cysteine, NADH* and glycerate-3-phosphate were obtained from Boehringer Mannheim. Trypsin and Dowex 50 WX2 were from Serva Feinbiochemica. BisCorrespondence to K. Bauer, Max-Planck-Institut fur experimentelle Endokrinologie, Feodor-Lynen-StraBe 7, D-30625 Hannover, Germany benzimide H 33258 fluorochrome was purchased from Calbiochem-Nova-biochem. Scintillator Hydroluma (J. T. Baker) was used for scintillation counting. Glass-fiber filters GF51 were obtained from Schleicher & Schuell. All solvents used were from Merck. Embryo extract was prepared according to the procedure of Konigsberg [8]. Cell culturePrimary cultures of myoblasts were isolated from the pectoral muscle of ll-day-old chick embryos. The muscle tissue obtained from 10 embryos was digested with 5 ml Puck's saline D2 solution (137 mM NaCl, 5.5 mM KCI, 0.22 mM KH2P04, 0.168 mM Na,HPO,, 0.11 mM CaCl, and 5.5 mM glucose) containing ...

show abstract

Section: Discussionsupporting

confidence: 82%

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

Bakardjiev¹,

Bauer²

1994

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…H'-coupled /I-alanine transport was inhibited by L-alanine [14] and L-proline (unpublished observation). In contrast, studies in rabbit intestinal BBMV demonstrated that NaCl-dependent j?-alanine transport was unaffected by L-alanine or Lproline [20] and p-alanine is not an inhibitor of Na'-dependent proline transport by the Imino carrier [5]. Although proline was a potent inhibitor of H'-coupled B-alanine transport and accumulation the question of whether these two substrates are transported by a single H'-coupled carrier is a difficult one to approach and remains to be answered.…”

Section: Discussionmentioning

confidence: 84%

H⁺‐coupled (Na⁺‐independent) proline transport in human intestinal (Caco‐2) epithelial cell monolayers

et al. 1993

View full text Add to dashboard Cite

Previously, absorption of l‐proline across the apical membrane of the intestinal enterocyte has been attributed to transport via the Na+‐dependent Imino system. However, net (absorptive) transport of proline across intact Caco‐2 cell monolayers was enhanced by acidification of the apical environment, under both Na+‐containing and Na+‐free conditions. This Na+‐independent pH‐dependent proline flux was associated with H+ flow across the apical membrane as determined by continuous measurement of intracellular pH. H+/proline symport was associated with an inward I sc in voltage‐clamped Caco‐2 epithelial layers demonstrating the electrogenic nature of this transport process. In conclusion Caco‐2 cells possess an apically‐localised, Na+‐independent, electrogenic H+/imino acid transporter which may play an important role in intestinal proline absorption.

show abstract

“…Transport is dependent upon Na + and Cl ¡ in the ratio 2:1:1 (Na + :Cl ¡ : -alanine) (Miyamoto et al 1990;Ramamoorthy et al 1994;Bakardjiev and Bauer 1994;Komura et al 1996). -alanine transport exhibits Michaelis-Menten kinetics with a Km in chick pectoral muscle of approximately 40 M (Bakardjiev and Bauer 1994;Kalyankar and Meister 1959;Winnick and Winnick 1959).…”

Section: Discussionmentioning

confidence: 99%

The effect of 4 weeks β-alanine supplementation and isokinetic training on carnosine concentrations in type I and II human skeletal muscle fibres

et al. 2009

View full text Add to dashboard Cite

Seven male students were supplemented with beta-alanine (beta-ALG) for 4 weeks (6.4 g day(-1)) and seven with a matching placebo (PLG). Subjects undertook 4 weeks of isokinetic training with the right leg (T) whilst the left leg was untrained (UT), serving as a control. Each training session consisted of 10 x 10 maximal 90 degrees extension and flexion contractions at 180 degrees /s using a Kin-Com isokinetic dynamometer, with 1 min rest between bouts. Muscle biopsies were taken from the vastus lateralis immediately before and at the end of the supplementation period. Following freeze drying muscle fibres were dissected and characterised by their MHC profile, as type I, IIa, IIx, or as hybrids of these. Carnosine was measured by HPLC. There was a significant increase in carnosine in both T and UT legs of the beta-ALG (9.63 +/- 3.92 mmol kg(-1) dry muscle and 6.55 +/- 2.36 mmol kg(-1) dry muscle respectively). There was a significant increase in the carnosine content of all fibre phentotypes, with no significant difference between types. There were no significant differences in the changes in muscle or in fibres between the T and UT legs. In contrast there was no significant change in the carnosine content in either the T or UT legs with placebo. The results indicate that 4 weeks training has no effect on the muscle carnosine content. Whilst an increase was seen with beta-alanine supplementation, this was not further influenced by training. These findings suggest that beta-alanine availability is the main factor regulating muscle carnosine synthesis.

show abstract

Uphill transport of beta-alanine in intestinal brush-border membrane vesicles

Cited by 22 publications

References 0 publications

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

H⁺‐coupled (Na⁺‐independent) proline transport in human intestinal (Caco‐2) epithelial cell monolayers

The effect of 4 weeks β-alanine supplementation and isokinetic training on carnosine concentrations in type I and II human skeletal muscle fibres

Contact Info

Product

Resources

About

Uphill transport of beta-alanine in intestinal brush-border membrane vesicles

Cited by 22 publications

References 0 publications

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

Transport of β‐alanine and biosynthesis of carnosine by skeletal muscle cells in primary culture

H+‐coupled (Na+‐independent) proline transport in human intestinal (Caco‐2) epithelial cell monolayers

The effect of 4 weeks β-alanine supplementation and isokinetic training on carnosine concentrations in type I and II human skeletal muscle fibres

Contact Info

Product

Resources

About

H⁺‐coupled (Na⁺‐independent) proline transport in human intestinal (Caco‐2) epithelial cell monolayers