Adaptive regulation of brush-border amino acid transport in a chronic excluded jejunal limb

1991 ◽  
Vol 261 (1) ◽  
pp. G22-G27
Author(s):  
R. M. Salloum ◽  
B. R. Stevens ◽  
W. W. Souba

We examined the alterations in brush-border glutamine transport that occurred in a surgically defunctionalized jejunal limb excluded from mucosal food contact. Dogs were surgically prepared with Roux-en-Y gastrojejunostomies to permit same-intestine comparisons of glutamine transport and glutaminase activity in jejunal segments that were in incontinuity or excluded for a 6-mo period. Transport of glutamine, alanine, and glucose was measured in brush-border membrane vesicles prepared from each intestinal section; membrane marker enzymes were enriched to the same degree in incontinuity and excluded portions. The Na(+)-dependent glutamine cotransport apparent Km was the same in the excluded (779 +/- 63 microM) and incontinuity (873 +/- 105 microM) limbs. However, the Jmax for Na(+)-independent glutamine transport in the incontinuity jejunum (158.7 +/- 15.7 pmol.mg protein-1.s-1) was double that in the excluded limb (71.2 +/- 4.6 pmol.mg protein-1.s-1). Na(+)-dependent carrier-mediated glutamine transport rates were lower than the Na(+)-dependent system, but Na(+)-independent kinetic parameters were not significantly different in incontinuity vs. excluded limbs (Jmax 7.9 +/- 0.6 pmol.mg protein-1.s-1; Km 140 +/- 20 microM). Similarly, the passive diffusion permeability coefficient was the same for both excluded and incontinuity jejunal limbs (22.7 +/- 0.9 nl.mg protein-1.s-1). Mucosal glutaminase enzyme activity was increased by 28% in the incontinuity limb (4.32 +/- 0.21 vs. 3.36 +/- 0.35 mumol.mg protein-1.h-1; P less than 0.02). Transport rates of alanine and glucose were also diminished in the excluded limb (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

1993 ◽  
Vol 290 (1) ◽  
pp. 237-240 ◽  
Author(s):  
H M Said ◽  
R Mohammadkhani

We examined the possible existence of histidine residue(s) in the folate transporter of rabbit intestine. This was done with use of the histidine-specific reagent diethyl pyrocarbonate (DEPC) and purified intestinal brush-border-membrane vesicles. DEPC caused significant concentration- and time-dependent inhibition of folic acid transport. The inhibition was only seen when transport was examined in vesicles incubated in buffer at pH 5.2 and not in those incubated in buffer at pH 7.4. The addition of unlabelled folic acid to vesicle suspension before treatment with DEPC (2.5 mM) led to a significant (P < 0.01) protection (84%) against the inhibition of folic acid transport. Treating vesicles pretreated with DEPC (2.5 mM) with reducing reagents (dithiothreitol, 2-mercaptoethanol and 2,3-dimercaptopropanol, all at a final concentration of 10 mM) did not reverse the inhibitory effect of DEPC on folic acid transport. On the other hand, treating the DEPC-pretreated vesicles with hydroxylamine (140 mM) led to a significant reversal (P < 0.01) (54%) of the inhibition of folic acid transport. The inhibitory effect of DEPC on carrier-mediated folic acid transport was found to be mediated through a decrease in the Vmax. (i.e. a decrease in the number and/or activity) of the carriers and an increase in the apparent Km (i.e. a decrease in their affinity), classifying the effect as a mixed-type inhibition. These results demonstrate the existence of critical histidine residue(s) in the intestinal brush-border-membrane folate transporter which is essential for its interaction with, and transport of, the vitamin. These findings also suggest that the histidine residue(s) is located at (or near) the substrate-binding site.


1986 ◽  
Vol 90 (5) ◽  
pp. 1188-1196 ◽  
Author(s):  
M.Susan Moyer ◽  
James E. Heubi ◽  
Anita L. Goodrich ◽  
William F. Balistreri ◽  
Frederick J. Suchy

1990 ◽  
Vol 27 (5) ◽  
pp. 519-524 ◽  
Author(s):  
Fadheela T Al-Mahroos ◽  
Nada Bulus ◽  
Naji Abumrad ◽  
Hamid Said ◽  
Fayez K Ghishan

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