Localization of the insulin-like growth factor II (IGF-II) binding/cross- linking site of the IGF-II/mannose 6-phosphate receptor to extracellular repeats 10-11

Farideh Garmroudi, Richard G MacDonald

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The insulin-like growth factor II (IGF-II) binding/cross-linking domain of the IGF-II/Man-6-P receptor was mapped by sequencing receptor fragments covalently attached to IGF-II. Rat placental or bovine liver receptors were purified by pentamannosyl-6-phosphate-Sepharose chromatography, affinity- labeled with 125I-IGF-II using disuccinimidyl tartrate, and digested with endoproteinase Glu-C. Analysis of small scale digests by gel electrophoresis revealed radiolabeled bands of ~17 kDa (rat) or ~18 kDa (bovine). For purification and sequencing of these radiolabeled receptor fragments, three receptor preparations were analyzed. The initial digests were fractionated by gel filtration followed by reverse-phase high performance liquid chromatography (HPLC), but the final purification steps differed somewhat in the three studies, using combinations of two-dimensional HPLC, gel electrophoresis, and electroblotting. Multiple sequences detected in each of these samples were unscrambled by computer-assisted and manual methods and by comparison with the quantity of labeled IGF-II present to identify sequences corresponding to fragments of the receptor covalently attached to IGF-II. The sequence, S(H)VNSXPMF, located in the COOH-terminal end of extracellular repeat 10 and beginning with serine 1488 of the bovine receptor, was the only receptor sequence common to all the samples and was the best candidate for the IGF-II cross-linked peptide by quantitative analysis. These data indicate residues within repeats 10-11 are likely to be important for IGF-II binding. We conclude that cross-linking between IGF-II and its receptor involves one or more of the 4 lysine residues located within extracellular repeat 11.

Original languageEnglish (US)
Pages (from-to)26944-26952
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number43
StatePublished - Jan 1 1994

Fingerprint

IGF Type 2 Receptor
Insulin-Like Growth Factor II
Crosslinking
Gels
High performance liquid chromatography
Electrophoresis
Purification
Rats
High Pressure Liquid Chromatography
Affinity chromatography
Agarose Chromatography
Reverse-Phase Chromatography
Liver
Sepharose
Serine
Lysine
Gel Chromatography
Phosphates
Peptides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{9c1e6ea5bcca45499fd7a6d1fe5a8bba,
title = "Localization of the insulin-like growth factor II (IGF-II) binding/cross- linking site of the IGF-II/mannose 6-phosphate receptor to extracellular repeats 10-11",
abstract = "The insulin-like growth factor II (IGF-II) binding/cross-linking domain of the IGF-II/Man-6-P receptor was mapped by sequencing receptor fragments covalently attached to IGF-II. Rat placental or bovine liver receptors were purified by pentamannosyl-6-phosphate-Sepharose chromatography, affinity- labeled with 125I-IGF-II using disuccinimidyl tartrate, and digested with endoproteinase Glu-C. Analysis of small scale digests by gel electrophoresis revealed radiolabeled bands of ~17 kDa (rat) or ~18 kDa (bovine). For purification and sequencing of these radiolabeled receptor fragments, three receptor preparations were analyzed. The initial digests were fractionated by gel filtration followed by reverse-phase high performance liquid chromatography (HPLC), but the final purification steps differed somewhat in the three studies, using combinations of two-dimensional HPLC, gel electrophoresis, and electroblotting. Multiple sequences detected in each of these samples were unscrambled by computer-assisted and manual methods and by comparison with the quantity of labeled IGF-II present to identify sequences corresponding to fragments of the receptor covalently attached to IGF-II. The sequence, S(H)VNSXPMF, located in the COOH-terminal end of extracellular repeat 10 and beginning with serine 1488 of the bovine receptor, was the only receptor sequence common to all the samples and was the best candidate for the IGF-II cross-linked peptide by quantitative analysis. These data indicate residues within repeats 10-11 are likely to be important for IGF-II binding. We conclude that cross-linking between IGF-II and its receptor involves one or more of the 4 lysine residues located within extracellular repeat 11.",
author = "Farideh Garmroudi and MacDonald, {Richard G}",
year = "1994",
month = "1",
day = "1",
language = "English (US)",
volume = "269",
pages = "26944--26952",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "43",

}

TY - JOUR

T1 - Localization of the insulin-like growth factor II (IGF-II) binding/cross- linking site of the IGF-II/mannose 6-phosphate receptor to extracellular repeats 10-11

AU - Garmroudi, Farideh

AU - MacDonald, Richard G

PY - 1994/1/1

Y1 - 1994/1/1

N2 - The insulin-like growth factor II (IGF-II) binding/cross-linking domain of the IGF-II/Man-6-P receptor was mapped by sequencing receptor fragments covalently attached to IGF-II. Rat placental or bovine liver receptors were purified by pentamannosyl-6-phosphate-Sepharose chromatography, affinity- labeled with 125I-IGF-II using disuccinimidyl tartrate, and digested with endoproteinase Glu-C. Analysis of small scale digests by gel electrophoresis revealed radiolabeled bands of ~17 kDa (rat) or ~18 kDa (bovine). For purification and sequencing of these radiolabeled receptor fragments, three receptor preparations were analyzed. The initial digests were fractionated by gel filtration followed by reverse-phase high performance liquid chromatography (HPLC), but the final purification steps differed somewhat in the three studies, using combinations of two-dimensional HPLC, gel electrophoresis, and electroblotting. Multiple sequences detected in each of these samples were unscrambled by computer-assisted and manual methods and by comparison with the quantity of labeled IGF-II present to identify sequences corresponding to fragments of the receptor covalently attached to IGF-II. The sequence, S(H)VNSXPMF, located in the COOH-terminal end of extracellular repeat 10 and beginning with serine 1488 of the bovine receptor, was the only receptor sequence common to all the samples and was the best candidate for the IGF-II cross-linked peptide by quantitative analysis. These data indicate residues within repeats 10-11 are likely to be important for IGF-II binding. We conclude that cross-linking between IGF-II and its receptor involves one or more of the 4 lysine residues located within extracellular repeat 11.

AB - The insulin-like growth factor II (IGF-II) binding/cross-linking domain of the IGF-II/Man-6-P receptor was mapped by sequencing receptor fragments covalently attached to IGF-II. Rat placental or bovine liver receptors were purified by pentamannosyl-6-phosphate-Sepharose chromatography, affinity- labeled with 125I-IGF-II using disuccinimidyl tartrate, and digested with endoproteinase Glu-C. Analysis of small scale digests by gel electrophoresis revealed radiolabeled bands of ~17 kDa (rat) or ~18 kDa (bovine). For purification and sequencing of these radiolabeled receptor fragments, three receptor preparations were analyzed. The initial digests were fractionated by gel filtration followed by reverse-phase high performance liquid chromatography (HPLC), but the final purification steps differed somewhat in the three studies, using combinations of two-dimensional HPLC, gel electrophoresis, and electroblotting. Multiple sequences detected in each of these samples were unscrambled by computer-assisted and manual methods and by comparison with the quantity of labeled IGF-II present to identify sequences corresponding to fragments of the receptor covalently attached to IGF-II. The sequence, S(H)VNSXPMF, located in the COOH-terminal end of extracellular repeat 10 and beginning with serine 1488 of the bovine receptor, was the only receptor sequence common to all the samples and was the best candidate for the IGF-II cross-linked peptide by quantitative analysis. These data indicate residues within repeats 10-11 are likely to be important for IGF-II binding. We conclude that cross-linking between IGF-II and its receptor involves one or more of the 4 lysine residues located within extracellular repeat 11.

UR - http://www.scopus.com/inward/record.url?scp=0027997861&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027997861&partnerID=8YFLogxK

M3 - Article

C2 - 7929433

AN - SCOPUS:0027997861

VL - 269

SP - 26944

EP - 26952

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 43

ER -