Abstract
Glucagon-like peptide-2 (GLP-2) has recently been identified as a stimulator of intestinal epithelial growth, prompting the development of RIA and HPLC methodologies to study this peptide in more detail. A GLP-2-specific antiserum (UTTH-7) was developed that recognizes amino acids 25-30 of human and rat GLP-2-(1-33). UTTH-7 cross-reacts with N- and C-terminally modified forms of GLP-2, proglucagon, and the major proglucagon fragment. Analysis of rat ileal extracts demonstrated the presence of GLP-2-(1-33) as well as significant amounts of GLP-2-(3-33) (16 ± 7% of total GLP-2). The level of total immunoreactive GLP-2 in plasma from fasted rats was 700 ± 71 pg/ml, and this increased 3.6-fold (P <0.001) in 24-h fed rate. HPLC analysis demonstrated the presence of both GLP-2-(1-33) and GLP-2-(3-38) in plasma from fasted rats, with increments in both peptides in plasma from fed rats. Immunoreactive GLP-2 increased in plasma from human subjects 2 h after a meal, rising from 851 ± 230 to 1106 ± 211 pg/ml (P <0.05); 15 ± 4% of this immunoreactivity was accounted for by the presence of intact GLP-2. HPLC showed the presence of both GLP-2-(1-33) and GLP-2-(3-33) in plasma from fed humans. Incubation of human GLP-2-(1-33) with the enzyme dipeptidylpeptidase IV resulted in liberation of GLP-2-(3-33), whereas replacement of Ala 2 with Gly 2 prevented this cleavage. Thus, while GLP-2-(1-33) is a major circulating and tissue form of GLP-2. GLP-2-(3-33) is a significant component of immunoreactive GLP-2 in both intestine and plasma.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 4837-4843 |
| Number of pages | 7 |
| Journal | Endocrinology |
| Volume | 138 |
| Issue number | 11 |
| DOIs | |
| State | Published - 1997 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Endocrinology
- Endocrinology, Diabetes and Metabolism
Cite this
Circulating and tissue forms of the intestinal growth factor, glucagon- like peptide-2. / Brubaker, Patricia L.; Crivici, Anna; Izzo, Angelo; Ehrlich, Peter; Tsai, Anne; Drucker, Daniel J.
In: Endocrinology, Vol. 138, No. 11, 1997, p. 4837-4843.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Circulating and tissue forms of the intestinal growth factor, glucagon- like peptide-2
AU - Brubaker, Patricia L.
AU - Crivici, Anna
AU - Izzo, Angelo
AU - Ehrlich, Peter
AU - Tsai, Anne
AU - Drucker, Daniel J.
PY - 1997
Y1 - 1997
N2 - Glucagon-like peptide-2 (GLP-2) has recently been identified as a stimulator of intestinal epithelial growth, prompting the development of RIA and HPLC methodologies to study this peptide in more detail. A GLP-2-specific antiserum (UTTH-7) was developed that recognizes amino acids 25-30 of human and rat GLP-2-(1-33). UTTH-7 cross-reacts with N- and C-terminally modified forms of GLP-2, proglucagon, and the major proglucagon fragment. Analysis of rat ileal extracts demonstrated the presence of GLP-2-(1-33) as well as significant amounts of GLP-2-(3-33) (16 ± 7% of total GLP-2). The level of total immunoreactive GLP-2 in plasma from fasted rats was 700 ± 71 pg/ml, and this increased 3.6-fold (P <0.001) in 24-h fed rate. HPLC analysis demonstrated the presence of both GLP-2-(1-33) and GLP-2-(3-38) in plasma from fasted rats, with increments in both peptides in plasma from fed rats. Immunoreactive GLP-2 increased in plasma from human subjects 2 h after a meal, rising from 851 ± 230 to 1106 ± 211 pg/ml (P <0.05); 15 ± 4% of this immunoreactivity was accounted for by the presence of intact GLP-2. HPLC showed the presence of both GLP-2-(1-33) and GLP-2-(3-33) in plasma from fed humans. Incubation of human GLP-2-(1-33) with the enzyme dipeptidylpeptidase IV resulted in liberation of GLP-2-(3-33), whereas replacement of Ala 2 with Gly 2 prevented this cleavage. Thus, while GLP-2-(1-33) is a major circulating and tissue form of GLP-2. GLP-2-(3-33) is a significant component of immunoreactive GLP-2 in both intestine and plasma.
AB - Glucagon-like peptide-2 (GLP-2) has recently been identified as a stimulator of intestinal epithelial growth, prompting the development of RIA and HPLC methodologies to study this peptide in more detail. A GLP-2-specific antiserum (UTTH-7) was developed that recognizes amino acids 25-30 of human and rat GLP-2-(1-33). UTTH-7 cross-reacts with N- and C-terminally modified forms of GLP-2, proglucagon, and the major proglucagon fragment. Analysis of rat ileal extracts demonstrated the presence of GLP-2-(1-33) as well as significant amounts of GLP-2-(3-33) (16 ± 7% of total GLP-2). The level of total immunoreactive GLP-2 in plasma from fasted rats was 700 ± 71 pg/ml, and this increased 3.6-fold (P <0.001) in 24-h fed rate. HPLC analysis demonstrated the presence of both GLP-2-(1-33) and GLP-2-(3-38) in plasma from fasted rats, with increments in both peptides in plasma from fed rats. Immunoreactive GLP-2 increased in plasma from human subjects 2 h after a meal, rising from 851 ± 230 to 1106 ± 211 pg/ml (P <0.05); 15 ± 4% of this immunoreactivity was accounted for by the presence of intact GLP-2. HPLC showed the presence of both GLP-2-(1-33) and GLP-2-(3-33) in plasma from fed humans. Incubation of human GLP-2-(1-33) with the enzyme dipeptidylpeptidase IV resulted in liberation of GLP-2-(3-33), whereas replacement of Ala 2 with Gly 2 prevented this cleavage. Thus, while GLP-2-(1-33) is a major circulating and tissue form of GLP-2. GLP-2-(3-33) is a significant component of immunoreactive GLP-2 in both intestine and plasma.
UR - http://www.scopus.com/inward/record.url?scp=0030782159&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030782159&partnerID=8YFLogxK
U2 - 10.1210/en.138.11.4837
DO - 10.1210/en.138.11.4837
M3 - Article
C2 - 9348213
AN - SCOPUS:0030782159
VL - 138
SP - 4837
EP - 4843
JO - Endocrinology
JF - Endocrinology
SN - 0013-7227
IS - 11
ER -