Intestinal brush border aminooligopeptidases: cytosol precursors of the membrane enzyme

Maze, Mervyn; Gray, Gary M.

doi:10.1021/bi00552a011

Cited by 32 publications

(10 citation statements)

References 29 publications

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“…Once obtained, these antibodies were subsequently coupled to Affi 10 agarose particles, which contain a linker molecule activated with an N ‐hydroxysuccinimide ester moiety. Upon reaction with a primary alkyl or aryl amine group on an amino acid residue, these linker molecules serve to couple the immunoglobulin to the stationary chromatographic phase via a hydrolytically stable amide bond 11. Once formed, the antibody‐containing agarose particles can be used to perform affinity purification of whole cell lysate of B. anthracis .…”

Section: Resultsmentioning

confidence: 99%

Simultaneous identification and verification of Bacillus anthracis

Krishnamurthy

Hewel

Bonzagni

et al. 2006

Rapid Comm Mass Spectrometry

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Specific identification of Bacillus anthracis (B. anthracis) is vital for the accurate treatment of afflicted personnel during biological warfare situations and civilian terrorist attacks. In order to accomplish this, we have subjected the lysates from B. anthracis to affinity purification using monoclonal antibodies for the selected antigenic protein present in the bacteria. The bound antigenic protein was identified by multi-dimensional protein identification technology (MudPIT) to be a surface layer protein EA1. The same antigen was identified from the lysates from a few strains of B. anthracis demonstrating the observation to be common for B. anthracis strains. Hence, this presents an effective pathway for the identification of the bacteria present in unknown samples of various origins. Generation of a database containing the EA1 protein has been found to be useful in the database search of unknown samples.

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Section: Resultsmentioning

confidence: 99%

Simultaneous identification and verification of Bacillus anthracis

Krishnamurthy

Hewel

Bonzagni

et al. 2006

Rapid Comm Mass Spectrometry

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“…It should be noted that soluble forms of brush-border membrane enzymes have already been reported in large amounts in the neonatal small intestine [11,39,41] and during organ culture of the small intestine [3]. They have also been characterized in intestinal tissues and found to present some differences with their brush-border counterparts [4,25]. However, their origin and role are only speculative [3,4,25,41].…”

Section: Discussionmentioning

confidence: 99%

“…They have also been characterized in intestinal tissues and found to present some differences with their brush-border counterparts [4,25]. However, their origin and role are only speculative [3,4,25,41].…”

Section: Discussionmentioning

confidence: 99%

Common characteristics for Na+-dependent sugar transport in Caco-2 cells and human fetal colon

1987

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The recent demonstration that the human colon adenocarcinoma cell line Caco-2 was susceptible to spontaneous enterocytic differentiation led us to consider the question as to whether Caco-2 cells would exhibit sodium-coupled transport of sugars. This problem was investigated using isotopic tracer flux measurements of the nonmetabolizable sugar analog alpha-methylglucoside (AMG). AMG accumulation in confluent monolayers was inhibited to the same extent by sodium replacement, 200 microM phlorizin, 1 mM phloretin, and 25 mM D-glucose, but was not inhibited further in the presence of both phlorizin and phloretin. Kinetic studies were compatible with the presence of both a simple diffusive process and a single, Na+-dependent, phlorizin- and phloretin-sensitive AMG transport system. These results also ruled out any interaction between AMG and a Na+-independent, phloretin-sensitive, facilitated diffusion pathway. The brush-border membrane localization of the Na+-dependent system was inferred from the observations that its functional differentiation was synchronous with the development of brush-border membrane enzyme activities and that phlorizin and phloretin addition 1 hr after initiating sugar transport produced immediate inhibition of AMG uptake as compared to ouabain. Finally, it was shown that brush-border membrane vesicles isolated from the human fetal colonic mucosa do possess a Na+-dependent transport pathway(s) for D-glucose which was inhibited by AMG and both phlorizin and phloretin. Caco-2 cells thus appear as a valuable cell culture model to study the mechanisms involved in the differentiation and regulation of intestinal transport functions.

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“…In the case of eukaryotic cells, the principal translocationcompetent membrane (for non-mitochondrial proteins) is the rough endoplasmic reticulum. This organelle, as mentioned above, is believed to function in a co-translational manner, but the surprising results of Cezard et al (1979) and Maze & Gray (1980), reporting the existence of soluble precursor forms of rat sucrase-isomaltase and aminopeptidase N, argued in favour of a post-translational membrane insertion.…”

Section: Introductionmentioning

confidence: 97%