Rat epidermal growth factor

complete amino acid sequence: Homology with the correspondence murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity

Richard J. SIMPSON, John A. SMITH, Robert L. MORITZ, Michael J. O'HARE, Philip S. RUDLAND, John R. MORRISON, Christopher J. LLOYD, Boris GREGO, Antony W. BURGESS, Edouard C. NICE

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Abstract

Epidermal growth factor (EGF) isolated from the submaxillary gland of the rat (rEGF) is missing the COOH‐terminal five residues present in both mouse and human EGF. rEGF competes for the binding of 125I‐labelled mEGF to human carcinoma cells with the same affinity as mEGF. rEGF and mEGF have identical mitogenic activities on mouse 3T3 fibroblasts, thus the C‐terminal region of the sequence is not necessary for the in vitro activity of EGF. Using reversed‐phase high‐performance liquid chromatography, four molecular forms of EGF have been extracted from rat submaxillary glands. These forms represent rEGF, rEGF(2–48), rEGF(3–48) and rEGF(4–48); all forms appear to be equipotent in both the receptor binding and mitogenic assays. The isoelectric points of these rEGFs are in the range of pH 5.1 to 5.2. The primary structure of rEGF was determined from approximately 10μg protein by sequence analysis of the intact molecule and fragments obtained from the reduced and alkylated protein by chemical cleavage with CNBr and enzymic cleavage with chymotrypsin and a proline‐specific endopeptidase. Subnanomole amounts of generated peptides were purified to homogeneity by reversed‐phase microbore high‐performance liquid chromatography and analysed by automated Edman degradation in a gas‐phase sequencer. There are 48 amino acid residues in the complete polypeptide chain which lacks alanine, phenylalanine, lysine and tryptophan. The amino acid sequence of rat epidermal growth factor is: Asn‐Ser‐Asn‐Thr‐ Gly‐Cys‐Pro‐Pro‐Ser‐Tyr‐Asp‐Gly‐Tyr‐Cys‐Leu‐Asn‐Gly‐Gly‐Val‐Cys‐Met‐Tyr‐Val‐Glu‐Ser‐Val‐Asp‐Arg‐Tyr‐Val‐Cys‐Asn‐Cys‐Val‐Ile‐Gly‐Tyr‐Ile‐Gly‐Glu‐Arg‐Cys‐Gln‐His‐Arg‐Leu‐Arg. The calculated relative molecular mass from the sequence analysis is 5377.

Original languageEnglish
Pages (from-to)629-637
Number of pages9
JournalEuropean Journal of Biochemistry
Volume153
Issue number3
DOIs
Publication statusPublished - 1 Jan 1985
Externally publishedYes

Cite this

SIMPSON, Richard J. ; SMITH, John A. ; MORITZ, Robert L. ; O'HARE, Michael J. ; RUDLAND, Philip S. ; MORRISON, John R. ; LLOYD, Christopher J. ; GREGO, Boris ; BURGESS, Antony W. ; NICE, Edouard C. / Rat epidermal growth factor : complete amino acid sequence: Homology with the correspondence murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity. In: European Journal of Biochemistry. 1985 ; Vol. 153, No. 3. pp. 629-637.
@article{e2261e5051f0436da6ef395aab6f3241,
title = "Rat epidermal growth factor: complete amino acid sequence: Homology with the correspondence murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity",
abstract = "Epidermal growth factor (EGF) isolated from the submaxillary gland of the rat (rEGF) is missing the COOH‐terminal five residues present in both mouse and human EGF. rEGF competes for the binding of 125I‐labelled mEGF to human carcinoma cells with the same affinity as mEGF. rEGF and mEGF have identical mitogenic activities on mouse 3T3 fibroblasts, thus the C‐terminal region of the sequence is not necessary for the in vitro activity of EGF. Using reversed‐phase high‐performance liquid chromatography, four molecular forms of EGF have been extracted from rat submaxillary glands. These forms represent rEGF, rEGF(2–48), rEGF(3–48) and rEGF(4–48); all forms appear to be equipotent in both the receptor binding and mitogenic assays. The isoelectric points of these rEGFs are in the range of pH 5.1 to 5.2. The primary structure of rEGF was determined from approximately 10μg protein by sequence analysis of the intact molecule and fragments obtained from the reduced and alkylated protein by chemical cleavage with CNBr and enzymic cleavage with chymotrypsin and a proline‐specific endopeptidase. Subnanomole amounts of generated peptides were purified to homogeneity by reversed‐phase microbore high‐performance liquid chromatography and analysed by automated Edman degradation in a gas‐phase sequencer. There are 48 amino acid residues in the complete polypeptide chain which lacks alanine, phenylalanine, lysine and tryptophan. The amino acid sequence of rat epidermal growth factor is: Asn‐Ser‐Asn‐Thr‐ Gly‐Cys‐Pro‐Pro‐Ser‐Tyr‐Asp‐Gly‐Tyr‐Cys‐Leu‐Asn‐Gly‐Gly‐Val‐Cys‐Met‐Tyr‐Val‐Glu‐Ser‐Val‐Asp‐Arg‐Tyr‐Val‐Cys‐Asn‐Cys‐Val‐Ile‐Gly‐Tyr‐Ile‐Gly‐Glu‐Arg‐Cys‐Gln‐His‐Arg‐Leu‐Arg. The calculated relative molecular mass from the sequence analysis is 5377.",
author = "SIMPSON, {Richard J.} and SMITH, {John A.} and MORITZ, {Robert L.} and O'HARE, {Michael J.} and RUDLAND, {Philip S.} and MORRISON, {John R.} and LLOYD, {Christopher J.} and Boris GREGO and BURGESS, {Antony W.} and NICE, {Edouard C.}",
year = "1985",
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language = "English",
volume = "153",
pages = "629--637",
journal = "European Journal of Biochemistry",
issn = "0014-2956",
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Rat epidermal growth factor : complete amino acid sequence: Homology with the correspondence murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity. / SIMPSON, Richard J.; SMITH, John A.; MORITZ, Robert L.; O'HARE, Michael J.; RUDLAND, Philip S.; MORRISON, John R.; LLOYD, Christopher J.; GREGO, Boris; BURGESS, Antony W.; NICE, Edouard C.

In: European Journal of Biochemistry, Vol. 153, No. 3, 01.01.1985, p. 629-637.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Rat epidermal growth factor

T2 - complete amino acid sequence: Homology with the correspondence murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity

AU - SIMPSON, Richard J.

AU - SMITH, John A.

AU - MORITZ, Robert L.

AU - O'HARE, Michael J.

AU - RUDLAND, Philip S.

AU - MORRISON, John R.

AU - LLOYD, Christopher J.

AU - GREGO, Boris

AU - BURGESS, Antony W.

AU - NICE, Edouard C.

PY - 1985/1/1

Y1 - 1985/1/1

N2 - Epidermal growth factor (EGF) isolated from the submaxillary gland of the rat (rEGF) is missing the COOH‐terminal five residues present in both mouse and human EGF. rEGF competes for the binding of 125I‐labelled mEGF to human carcinoma cells with the same affinity as mEGF. rEGF and mEGF have identical mitogenic activities on mouse 3T3 fibroblasts, thus the C‐terminal region of the sequence is not necessary for the in vitro activity of EGF. Using reversed‐phase high‐performance liquid chromatography, four molecular forms of EGF have been extracted from rat submaxillary glands. These forms represent rEGF, rEGF(2–48), rEGF(3–48) and rEGF(4–48); all forms appear to be equipotent in both the receptor binding and mitogenic assays. The isoelectric points of these rEGFs are in the range of pH 5.1 to 5.2. The primary structure of rEGF was determined from approximately 10μg protein by sequence analysis of the intact molecule and fragments obtained from the reduced and alkylated protein by chemical cleavage with CNBr and enzymic cleavage with chymotrypsin and a proline‐specific endopeptidase. Subnanomole amounts of generated peptides were purified to homogeneity by reversed‐phase microbore high‐performance liquid chromatography and analysed by automated Edman degradation in a gas‐phase sequencer. There are 48 amino acid residues in the complete polypeptide chain which lacks alanine, phenylalanine, lysine and tryptophan. The amino acid sequence of rat epidermal growth factor is: Asn‐Ser‐Asn‐Thr‐ Gly‐Cys‐Pro‐Pro‐Ser‐Tyr‐Asp‐Gly‐Tyr‐Cys‐Leu‐Asn‐Gly‐Gly‐Val‐Cys‐Met‐Tyr‐Val‐Glu‐Ser‐Val‐Asp‐Arg‐Tyr‐Val‐Cys‐Asn‐Cys‐Val‐Ile‐Gly‐Tyr‐Ile‐Gly‐Glu‐Arg‐Cys‐Gln‐His‐Arg‐Leu‐Arg. The calculated relative molecular mass from the sequence analysis is 5377.

AB - Epidermal growth factor (EGF) isolated from the submaxillary gland of the rat (rEGF) is missing the COOH‐terminal five residues present in both mouse and human EGF. rEGF competes for the binding of 125I‐labelled mEGF to human carcinoma cells with the same affinity as mEGF. rEGF and mEGF have identical mitogenic activities on mouse 3T3 fibroblasts, thus the C‐terminal region of the sequence is not necessary for the in vitro activity of EGF. Using reversed‐phase high‐performance liquid chromatography, four molecular forms of EGF have been extracted from rat submaxillary glands. These forms represent rEGF, rEGF(2–48), rEGF(3–48) and rEGF(4–48); all forms appear to be equipotent in both the receptor binding and mitogenic assays. The isoelectric points of these rEGFs are in the range of pH 5.1 to 5.2. The primary structure of rEGF was determined from approximately 10μg protein by sequence analysis of the intact molecule and fragments obtained from the reduced and alkylated protein by chemical cleavage with CNBr and enzymic cleavage with chymotrypsin and a proline‐specific endopeptidase. Subnanomole amounts of generated peptides were purified to homogeneity by reversed‐phase microbore high‐performance liquid chromatography and analysed by automated Edman degradation in a gas‐phase sequencer. There are 48 amino acid residues in the complete polypeptide chain which lacks alanine, phenylalanine, lysine and tryptophan. The amino acid sequence of rat epidermal growth factor is: Asn‐Ser‐Asn‐Thr‐ Gly‐Cys‐Pro‐Pro‐Ser‐Tyr‐Asp‐Gly‐Tyr‐Cys‐Leu‐Asn‐Gly‐Gly‐Val‐Cys‐Met‐Tyr‐Val‐Glu‐Ser‐Val‐Asp‐Arg‐Tyr‐Val‐Cys‐Asn‐Cys‐Val‐Ile‐Gly‐Tyr‐Ile‐Gly‐Glu‐Arg‐Cys‐Gln‐His‐Arg‐Leu‐Arg. The calculated relative molecular mass from the sequence analysis is 5377.

UR - http://www.scopus.com/inward/record.url?scp=0022296446&partnerID=8YFLogxK

U2 - 10.1111/j.1432-1033.1985.tb09346.x

DO - 10.1111/j.1432-1033.1985.tb09346.x

M3 - Article

VL - 153

SP - 629

EP - 637

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 3

ER -