Analysis of glycylsarcosine transport by lobster intestine using gas chromatography
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- 作者：Maria L. Peterson ; Amy L. Lane ; Gregory A. Ahearn
- 关键词：Homarus americanus ; Gas chromatography ; Intestinal transport ; Dipeptide ; Glycylsarcosine ; l ; Leucine ; Calcium ; Zinc ; Bis ; complex ; Molecular mimicry ; PEPT1 ; PHT1
- 刊名：Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology
- 出版年：2015
- 出版时间：January 2015
- 年：2015
- 卷：185
- 期：1
- 页码：37-45
- 全文大小：488 KB
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文摘
Gas chromatography was used to measure transepithelial transport of glycylsarcosine (Gly-Sar) by perfused lobster (Homarus americanus) intestine. Unidirectional and net fluxes of dipeptide across the tissue and luminal factors affecting their magnitude and direction were characterized by perfusing the lumen with the dipeptide and measuring its appearance in saline on the serosal side of the organ. Transmural transport of 10?mM Gly-Sar resulted in serosal accumulation of only the dipeptide; no appearance of corresponding monomeric amino acids glycine or sarcosine was observed. Carrier-mediated and diffusional transmural intestinal transport of Gly-Sar was estimated at 1-5?mM luminal concentrations and followed a curvilinear equation providing a K m?=?0.44?±?0.17?mM, a J max?=?1.27?±?0.12?nmol?cm??min?, and a diffusional coefficient?=?0.026?±?0.008?nmol?cm??min??mM?. Unidirectional mucosal to serosal and serosal to mucosal fluxes of 10?mM Gly-Sar provided a significant (p???min?, further supporting carrier-mediated dipeptide transport across the gut. Alkaline (pH 8.5) luminal pH more than doubled transmural Gly-Sar transport as compared to acidic (pH 5.5) luminal pH, while luminal amino acid-metal chelates (e.g., Leu-Zn-Leu), and high concentrations of amino acids alone significantly (p?