A possible high molecular weight precursor to vasoactive intestinal polypeptide sequestered into pheochromocytoma chromaffin granules

Illana Gozes; Daniel T. O'Connor; Floyd E. Bloom

doi:10.1016/0167-0115(83)90004-6

A possible high molecular weight precursor to vasoactive intestinal polypeptide sequestered into pheochromocytoma chromaffin granules

Illana Gozes, Daniel T. O'Connor, Floyd E. Bloom

Research output: Contribution to journal › Article › peer-review

Abstract

Chromaffin granules, the catecholamine storage granules of pheochromocytoma were isolated from five human pheochromocytoma tumors. Vasoactive intestinal polypeptide (VIP) immunoreactivity was detected in all chromaffin granule preparations, paralleling the synthetic VIP antibody binding curve over a range of serial dilutions. In addition, gel filtration revealed an immunoreactive peptide peak coeluting with VIP. However, high molecular weight immunoreactive material was also detected on the column. This high molecular weight material was further characterized by sodium dodecyl sulfate gel electrophoresis, followed by electroblotting onto nitrocellulose paper and detection by anti-VIP antibodies with a secondary antibody conjugated to horseradish peroxidase. A 70 000 dalton immunoreactive band was identified, in which reactivity with anti-VIP antibody was inhibited by VIP; this band did not cross react with non-related antibodies. This 70 000 dalton protein may be an intermediate molecule in the biosynthesis and processing of VIP.

Original language	English
Pages (from-to)	111-119
Number of pages	9
Journal	Regulatory Peptides
Volume	6
Issue number	2
DOIs	https://doi.org/10.1016/0167-0115(83)90004-6
State	Published - May 1983
Externally published	Yes

Keywords

VIP
chromaffin granules
electroblotting
gel electrophoresis
pheochromocytoma
radioimmunoassay

Access to Document

10.1016/0167-0115(83)90004-6

Cite this

@article{6e4cb95296884a659aaf02cfdab4020d,

title = "A possible high molecular weight precursor to vasoactive intestinal polypeptide sequestered into pheochromocytoma chromaffin granules",

abstract = "Chromaffin granules, the catecholamine storage granules of pheochromocytoma were isolated from five human pheochromocytoma tumors. Vasoactive intestinal polypeptide (VIP) immunoreactivity was detected in all chromaffin granule preparations, paralleling the synthetic VIP antibody binding curve over a range of serial dilutions. In addition, gel filtration revealed an immunoreactive peptide peak coeluting with VIP. However, high molecular weight immunoreactive material was also detected on the column. This high molecular weight material was further characterized by sodium dodecyl sulfate gel electrophoresis, followed by electroblotting onto nitrocellulose paper and detection by anti-VIP antibodies with a secondary antibody conjugated to horseradish peroxidase. A 70 000 dalton immunoreactive band was identified, in which reactivity with anti-VIP antibody was inhibited by VIP; this band did not cross react with non-related antibodies. This 70 000 dalton protein may be an intermediate molecule in the biosynthesis and processing of VIP.",

keywords = "VIP, chromaffin granules, electroblotting, gel electrophoresis, pheochromocytoma, radioimmunoassay",

author = "Illana Gozes and O'Connor, {Daniel T.} and Bloom, {Floyd E.}",

note = "Funding Information: We would like to thank Drs. Fahrenkrug and Benoit for the generous gift of anti-VIP antibodies and Dr. Gautsch for very helpful discussions. The excellent assistance of Ms. Raana Azad and Mr. Gary Wyatt is gratefully acknowledged. This work was supported by Del Webb Foundation Grants (to F.E.B.), The Veterans Administration, the National Institutes of Health (HL-25, 457), and the American Heart Association grants (to D.T.O.) and by a grant from the United States-Israel Binational Science Foundation (BSF) Jerusalem (to I,G. and F.E.B.), a 1982 Bergmann Memorial Research Grant (I.G.), a grant from The Fund for Basic Research Administered by the Israeli Academy of Sciences and Humanities (to I.G.), the Jewish Agency (I.G.) and the Leo and Julia Forchheimer Center for Molecular Genetics (I.G.).",

year = "1983",

month = may,

doi = "10.1016/0167-0115(83)90004-6",

language = "אנגלית",

volume = "6",

pages = "111--119",

journal = "Regulatory Peptides",

issn = "0167-0115",

publisher = "Elsevier",

number = "2",

}

TY - JOUR

T1 - A possible high molecular weight precursor to vasoactive intestinal polypeptide sequestered into pheochromocytoma chromaffin granules

AU - Gozes, Illana

AU - O'Connor, Daniel T.

AU - Bloom, Floyd E.

N1 - Funding Information: We would like to thank Drs. Fahrenkrug and Benoit for the generous gift of anti-VIP antibodies and Dr. Gautsch for very helpful discussions. The excellent assistance of Ms. Raana Azad and Mr. Gary Wyatt is gratefully acknowledged. This work was supported by Del Webb Foundation Grants (to F.E.B.), The Veterans Administration, the National Institutes of Health (HL-25, 457), and the American Heart Association grants (to D.T.O.) and by a grant from the United States-Israel Binational Science Foundation (BSF) Jerusalem (to I,G. and F.E.B.), a 1982 Bergmann Memorial Research Grant (I.G.), a grant from The Fund for Basic Research Administered by the Israeli Academy of Sciences and Humanities (to I.G.), the Jewish Agency (I.G.) and the Leo and Julia Forchheimer Center for Molecular Genetics (I.G.).

PY - 1983/5

Y1 - 1983/5

N2 - Chromaffin granules, the catecholamine storage granules of pheochromocytoma were isolated from five human pheochromocytoma tumors. Vasoactive intestinal polypeptide (VIP) immunoreactivity was detected in all chromaffin granule preparations, paralleling the synthetic VIP antibody binding curve over a range of serial dilutions. In addition, gel filtration revealed an immunoreactive peptide peak coeluting with VIP. However, high molecular weight immunoreactive material was also detected on the column. This high molecular weight material was further characterized by sodium dodecyl sulfate gel electrophoresis, followed by electroblotting onto nitrocellulose paper and detection by anti-VIP antibodies with a secondary antibody conjugated to horseradish peroxidase. A 70 000 dalton immunoreactive band was identified, in which reactivity with anti-VIP antibody was inhibited by VIP; this band did not cross react with non-related antibodies. This 70 000 dalton protein may be an intermediate molecule in the biosynthesis and processing of VIP.

AB - Chromaffin granules, the catecholamine storage granules of pheochromocytoma were isolated from five human pheochromocytoma tumors. Vasoactive intestinal polypeptide (VIP) immunoreactivity was detected in all chromaffin granule preparations, paralleling the synthetic VIP antibody binding curve over a range of serial dilutions. In addition, gel filtration revealed an immunoreactive peptide peak coeluting with VIP. However, high molecular weight immunoreactive material was also detected on the column. This high molecular weight material was further characterized by sodium dodecyl sulfate gel electrophoresis, followed by electroblotting onto nitrocellulose paper and detection by anti-VIP antibodies with a secondary antibody conjugated to horseradish peroxidase. A 70 000 dalton immunoreactive band was identified, in which reactivity with anti-VIP antibody was inhibited by VIP; this band did not cross react with non-related antibodies. This 70 000 dalton protein may be an intermediate molecule in the biosynthesis and processing of VIP.

KW - VIP

KW - chromaffin granules

KW - electroblotting

KW - gel electrophoresis

KW - pheochromocytoma

KW - radioimmunoassay

UR - http://www.scopus.com/inward/record.url?scp=0020960976&partnerID=8YFLogxK

U2 - 10.1016/0167-0115(83)90004-6

DO - 10.1016/0167-0115(83)90004-6

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

AN - SCOPUS:0020960976

VL - 6

SP - 111

EP - 119

JO - Regulatory Peptides

JF - Regulatory Peptides

SN - 0167-0115

IS - 2

ER -

A possible high molecular weight precursor to vasoactive intestinal polypeptide sequestered into pheochromocytoma chromaffin granules

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this