Characterization of immunoreactive motilin from the rat small intestine

1990 ◽  
Vol 68 (8) ◽  
pp. 1124-1130 ◽  
Author(s):  
L. B. Vogel ◽  
J. C. Brown
Keyword(s):  
Biological Activity ◽  
Small Intestine ◽  
Molecular Size ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
Muscle Strip ◽  
Rat Small Intestine ◽  
Fixation Techniques ◽  
Staining Methods

Immunocytochemistry, radioimmunoassay, chromatography, and biological assay using a rabbit isolated duodenal muscle strip preparation were used in attempting to characterize motilin from the rat small intestine. Several different antisera and monoclonal antibodies directed against natural porcine motilin were used. A variety of fixation techniques using Bouin's, paraformaldehyde, and benzoquinone with different staining methods including, fluorescein-conjugated second antibody, peroxidase–antiperoxidase or peroxidase-conjugated second antibody techniques were used. All methods failed to detect immunoreactive motilin cells in the rat small intestine. The same antisera were used in radioimmunoassays for motilin to evaluate extracts of rat intestinal tissue. Two of these detected immunoreactive motilin in gut extracts, and these antisera showed a different distribution for the peptide. Samples containing immunoreactive motilin obtained from cation exchange chromatography on SP-Sephadex-G25 were concentrated and assayed for biological activity in a rabbit duodenal muscle strip preparation. Desensitization of duodenal tissue to porcine motilin could be demonstrated by pretreatment with this peptide. The biological activity of partially purified rat intestinal immunoreactive motilin was not prevented by pretreatment of the tissue with motilin. Further purification of this preparation on Bio-Gel P-10 yielded an immunoreactive motilin peak that co-eluted with natural porcine motilin. Rat intestinal immunoreactive motilin did not co-elute with natural porcine motilin following high pressure liquid chromatography on a Waters μBondapak C18 reversed-phase column using a linear gradient of water–acetonitrile (10–45%) over 30 min. Although of similar molecular size, rat motilin is probably structurally dissimilar to other mammalian motilins.Key words: motilin, radioimmunoassay, immunocytochemistry.

Region-dependent disappearance of vinblastine in rat small intestine and characterization of its P-glycoprotein-mediated efflux system

2000 ◽  
Vol 11 (4) ◽  
pp. 317-324 ◽  
Author(s):  
Akira Nakayama ◽  
Hiroshi Saitoh ◽  
Masako Oda ◽  
Masahiko Takada ◽  
Bruce J Aungst
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine ◽  
Efflux System ◽  
P Glycoprotein

scholarly journals Characterization of aminocephalosporin transport across rat small intestine.

1983 ◽  
Vol 6 (4) ◽  
pp. 246-253 ◽  
Author(s):  
TOSHIKIRO KIMURA ◽  
TAKAO YAMAMOTO ◽  
MASAYUKI MIZUNO ◽  
YOSHIE SUGA ◽  
SUMIKO KITADE ◽  
...  
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine

S20.9 An insoluble mucin complex from rat small intestine ? Characterization of two different glycosylated domains

1993 ◽  
Vol 10 (4) ◽  
pp. 344-344 ◽  
Author(s):  
I. Carlstedt ◽  
A. Herrmann ◽  
H. Karlsson ◽  
J. K. Sheehan ◽  
L. -A. Fransson ◽  
...  
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine

Structural characterization of glycolipids of rat small intestine having one to eight hexoses in a linear sequence

1982 ◽  
Vol 213 (2) ◽  
pp. 708-725 ◽  
Author(s):  
Jonas Ångström ◽  
Michael E. Breimer ◽  
Karl-Erik Falk ◽  
Gunnar C. Hansson ◽  
Karl-Andres Karlsson ◽  
...  
Keyword(s):  
Small Intestine ◽  
Structural Characterization ◽  
Rat Small Intestine ◽  
Linear Sequence

scholarly journals Partial Chemical Characterization of S-Carboxymethylated Chains of Rabbit Fibrin(OGEN): Evidence for Separate Chain Biosynthesis

1977 ◽  
Author(s):  
B. Alving ◽  
G. Murano ◽  
D. Walz
Keyword(s):  
Amino Acid ◽  
Specific Activity ◽  
Chemical Characterization ◽  
Molecular Size ◽  
Exchange Chromatography ◽  
12 Steps ◽  
Polypeptide Chains ◽  
Sds Electrophoresis ◽  
Derivatives Of

The purpose of this study was twofold: 1) chemically characterize the isolated polypeptide chains of rabbit fibrin(ogen), and 2) explore their mode of biosynthesis. The three S-carboxy-methyl polypeptide chain derivatives of rabbit fibrin (α, β and γ) were isolated by cation exchange chromatography. Their amino acid composition was similar to the human with a methionine distribution (mole/mole) as follows: γ = 9; β = 14, α = 14. Their molecular size, (SDS electrophoresis) was estimated as follows: γ = 46,000; β = 54,000; α = 63,500. The N-terminal amino acid sequence (12 steps) of the β derivative was:Gly-His-Arg-Pro-Ile-Asp-Arg-Arg-Arg-Glu-Glu-Leu-. To determine whether the three chains are synthesized sequentially (one continuous chain, later split into three) or in parallel, turpentine-stimulated male New Zealand rabbits were given ~40 μCi of [75Se] selenomethionine (SeM) and its incorporation into fibrinogen (F) was followed. F was clotted from plasma samples, washed, reduced, and constituent chains separated by gel electrophoresis in the presence of SDS-urea. The radioactivity of each chain (expressed as percent of total F radioactivity) was determined, and the specific methionine radioactivity calculated for each chain isolated at 20, 25, and 30 min after SeM injection. During this interval the specific activity of the α and the γ chains was essentially the same (within 3%) while that of the β chain was 42 to 97% greater than that of the α chain. The similar activity of the α and γ chains during the early phase of SeM incorporation suggests that these two chains are not synthesized sequentially, rather they are synthesized in parallel.

Functional characterization of PCFT/HCP1 as the molecular entity of the carrier-mediated intestinal folate transport system in the rat model

2008 ◽  
Vol 294 (3) ◽  
pp. G660-G668 ◽  
Author(s):  
Katsuhisa Inoue ◽  
Yasuhiro Nakai ◽  
Sayaka Ueda ◽  
Shunsuke Kamigaso ◽  
Kin-ya Ohta ◽  
...  
Keyword(s):  
Small Intestine ◽  
Transport System ◽  
Functional Characterization ◽  
Cellular Membrane ◽  
Molecular Entity ◽  
Xenopus Laevis Oocytes ◽  
Rat Small Intestine ◽  
Distribution Profile ◽  
Folate Transport

Proton-coupled folate transporter/heme carrier protein 1 (PCFT/HCP1) has recently been identified as a transporter that mediates the translocation of folates across the cellular membrane by a proton-coupled mechanism and suggested to be the possible molecular entity of the carrier-mediated intestinal folate transport system. To further clarify its role in intestinal folate transport, we examined the functional characteristics of rat PCFT/HCP1 (rPCFT/HCP1) expressed in Xenopus laevis oocytes and compared with those of the carrier-mediated folate transport system in the rat small intestine evaluated by using the everted tissue sacs. rPCFT/HCP1 was demonstrated to transport folate and methotrexate more efficiently at lower acidic pH and, as evaluated at pH 5.5, with smaller Michaelis constant ( Km) for the former (2.4 μM) than for the latter (5.7 μM), indicating its characteristic as a proton-coupled folate transporter that favors folate than methotrexate as substrate. rPCFT/HCP1-mediated folate transport was found to be inhibited by several but limited anionic compounds, such as sulfobromophthalein and sulfasalazine. All these characteristics of rPCFT/HCP1 were in agreement with those of carrier-mediated intestinal folate transport system, of which the Km values were 1.2 and 5.8 μM for folate and methotrexate, respectively, in the rat small intestine. Furthermore, the distribution profile of the folate transport system activity along the intestinal tract was in agreement with that of rPCFT/HCP1 mRNA. This study is the first to clone rPCFT/HCP1, and we successfully provided several lines of evidence that indicate its role as the molecular entity of the intestinal folate transport system.

Characterization of WR-1065 absorption in the rat small intestine

1991 ◽  
Vol 12 (4) ◽  
pp. 275-284 ◽  
Author(s):  
R. S. Geary ◽  
N. F. Swynnerton ◽  
S. F. Timmons ◽  
D. J. Mangold
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine
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