Prouroguanylin and proguanylin: Purification from colon, structure, and modulation of bioactivity by proteases

F. Kent Hamra, Xiaohui Fan, William J. Krause, Ronald H. Freeman, David T. Chin, Christine E. Smith, Mark G. Currie, Leonard R. Forte

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Uroguanylin and guanylin are peptides isolated from urine and intestinal mucosa, which regulate cyclic GMP production in enterocytes by activating an apical membrane, receptor-guanylate cyclase. This study extended our previous findings, which showed that colonic mucosa of opossums contained uroguanylin and guanylin peptides, by purifying prouroguanylin and proguanylin from this tissue. Prouroguanylin and proguanylin coeluted from Sephadex G-75 gel- filtration columns with a similar molecular size between 6 and 12 kDa. Mass spectrometry indicated that proguanylin (≃8.7 kDa) had a 10% lower molecular mass than prouroguanylin (≃9.7 kDa). Isoelectric focusing separated prouroguanylin (pI ≃4.5) from proguanylin (pI ≃7.5). N-terminal sequence analysis of reverse phase-HPLC purified prohormones revealed 13 amino acids in opossum proguanylin that shared 77-85% identity with human and rat proguanylin, but only 23% identity with opossum prouroguanylin. The N- terminal 19 residues obtained for opossum prouroguanylin shared 32-42% identity with rat and human proguanylin. Prouroguanylin and proguanylin were both inactive and required pretreatment with proteases to elicit cyclic GMP responses in T84 cells. V8 protease treatment of proguanylin liberated a bioactive, 16-amino acid form of guanylin. Chymotrypsin treatment activated prouroguanylin, but inactivated the bioactive peptide domain within proguanylin. In summary, colonic mucosa contains the bioactive peptide and inactive prohormone forms of uroguanylin and guanylin. Thus, after proteolytic processing of prouroguanylin and proguanylin, bioactive uroguanylin and guanylin could both function to regulate guanylate cyclase activity by autocrine and/or paracrine actions on enterocytes. Also, these peptide hormones are implicated in an intestinal-renal axis for the endocrine regulation of salt and water homeostasis.

Original languageEnglish (US)
Pages (from-to)257-265
Number of pages9
JournalEndocrinology
Volume137
Issue number1
DOIs
StatePublished - 1996

Fingerprint

Colon
Peptide Hydrolases
Opossums
Enterocytes
Guanylate Cyclase
Cyclic GMP
Peptides
Mucous Membrane
prouroguanylin
preproguanylin
Amino Acids
Peptide Hormones
Chymotrypsin
Isoelectric Focusing
Intestinal Mucosa
Gel Chromatography
Sequence Analysis
Mass Spectrometry
Homeostasis
Salts

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Hamra, F. K., Fan, X., Krause, W. J., Freeman, R. H., Chin, D. T., Smith, C. E., ... Forte, L. R. (1996). Prouroguanylin and proguanylin: Purification from colon, structure, and modulation of bioactivity by proteases. Endocrinology, 137(1), 257-265. https://doi.org/10.1210/en.137.1.257

Prouroguanylin and proguanylin : Purification from colon, structure, and modulation of bioactivity by proteases. / Hamra, F. Kent; Fan, Xiaohui; Krause, William J.; Freeman, Ronald H.; Chin, David T.; Smith, Christine E.; Currie, Mark G.; Forte, Leonard R.

In: Endocrinology, Vol. 137, No. 1, 1996, p. 257-265.

Research output: Contribution to journalArticle

Hamra, FK, Fan, X, Krause, WJ, Freeman, RH, Chin, DT, Smith, CE, Currie, MG & Forte, LR 1996, 'Prouroguanylin and proguanylin: Purification from colon, structure, and modulation of bioactivity by proteases', Endocrinology, vol. 137, no. 1, pp. 257-265. https://doi.org/10.1210/en.137.1.257
Hamra, F. Kent ; Fan, Xiaohui ; Krause, William J. ; Freeman, Ronald H. ; Chin, David T. ; Smith, Christine E. ; Currie, Mark G. ; Forte, Leonard R. / Prouroguanylin and proguanylin : Purification from colon, structure, and modulation of bioactivity by proteases. In: Endocrinology. 1996 ; Vol. 137, No. 1. pp. 257-265.
@article{79505148f50b404aafe75d1ae4cbfd54,
title = "Prouroguanylin and proguanylin: Purification from colon, structure, and modulation of bioactivity by proteases",
abstract = "Uroguanylin and guanylin are peptides isolated from urine and intestinal mucosa, which regulate cyclic GMP production in enterocytes by activating an apical membrane, receptor-guanylate cyclase. This study extended our previous findings, which showed that colonic mucosa of opossums contained uroguanylin and guanylin peptides, by purifying prouroguanylin and proguanylin from this tissue. Prouroguanylin and proguanylin coeluted from Sephadex G-75 gel- filtration columns with a similar molecular size between 6 and 12 kDa. Mass spectrometry indicated that proguanylin (≃8.7 kDa) had a 10{\%} lower molecular mass than prouroguanylin (≃9.7 kDa). Isoelectric focusing separated prouroguanylin (pI ≃4.5) from proguanylin (pI ≃7.5). N-terminal sequence analysis of reverse phase-HPLC purified prohormones revealed 13 amino acids in opossum proguanylin that shared 77-85{\%} identity with human and rat proguanylin, but only 23{\%} identity with opossum prouroguanylin. The N- terminal 19 residues obtained for opossum prouroguanylin shared 32-42{\%} identity with rat and human proguanylin. Prouroguanylin and proguanylin were both inactive and required pretreatment with proteases to elicit cyclic GMP responses in T84 cells. V8 protease treatment of proguanylin liberated a bioactive, 16-amino acid form of guanylin. Chymotrypsin treatment activated prouroguanylin, but inactivated the bioactive peptide domain within proguanylin. In summary, colonic mucosa contains the bioactive peptide and inactive prohormone forms of uroguanylin and guanylin. Thus, after proteolytic processing of prouroguanylin and proguanylin, bioactive uroguanylin and guanylin could both function to regulate guanylate cyclase activity by autocrine and/or paracrine actions on enterocytes. Also, these peptide hormones are implicated in an intestinal-renal axis for the endocrine regulation of salt and water homeostasis.",
author = "Hamra, {F. Kent} and Xiaohui Fan and Krause, {William J.} and Freeman, {Ronald H.} and Chin, {David T.} and Smith, {Christine E.} and Currie, {Mark G.} and Forte, {Leonard R.}",
year = "1996",
doi = "10.1210/en.137.1.257",
language = "English (US)",
volume = "137",
pages = "257--265",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Prouroguanylin and proguanylin

T2 - Purification from colon, structure, and modulation of bioactivity by proteases

AU - Hamra, F. Kent

AU - Fan, Xiaohui

AU - Krause, William J.

AU - Freeman, Ronald H.

AU - Chin, David T.

AU - Smith, Christine E.

AU - Currie, Mark G.

AU - Forte, Leonard R.

PY - 1996

Y1 - 1996

N2 - Uroguanylin and guanylin are peptides isolated from urine and intestinal mucosa, which regulate cyclic GMP production in enterocytes by activating an apical membrane, receptor-guanylate cyclase. This study extended our previous findings, which showed that colonic mucosa of opossums contained uroguanylin and guanylin peptides, by purifying prouroguanylin and proguanylin from this tissue. Prouroguanylin and proguanylin coeluted from Sephadex G-75 gel- filtration columns with a similar molecular size between 6 and 12 kDa. Mass spectrometry indicated that proguanylin (≃8.7 kDa) had a 10% lower molecular mass than prouroguanylin (≃9.7 kDa). Isoelectric focusing separated prouroguanylin (pI ≃4.5) from proguanylin (pI ≃7.5). N-terminal sequence analysis of reverse phase-HPLC purified prohormones revealed 13 amino acids in opossum proguanylin that shared 77-85% identity with human and rat proguanylin, but only 23% identity with opossum prouroguanylin. The N- terminal 19 residues obtained for opossum prouroguanylin shared 32-42% identity with rat and human proguanylin. Prouroguanylin and proguanylin were both inactive and required pretreatment with proteases to elicit cyclic GMP responses in T84 cells. V8 protease treatment of proguanylin liberated a bioactive, 16-amino acid form of guanylin. Chymotrypsin treatment activated prouroguanylin, but inactivated the bioactive peptide domain within proguanylin. In summary, colonic mucosa contains the bioactive peptide and inactive prohormone forms of uroguanylin and guanylin. Thus, after proteolytic processing of prouroguanylin and proguanylin, bioactive uroguanylin and guanylin could both function to regulate guanylate cyclase activity by autocrine and/or paracrine actions on enterocytes. Also, these peptide hormones are implicated in an intestinal-renal axis for the endocrine regulation of salt and water homeostasis.

AB - Uroguanylin and guanylin are peptides isolated from urine and intestinal mucosa, which regulate cyclic GMP production in enterocytes by activating an apical membrane, receptor-guanylate cyclase. This study extended our previous findings, which showed that colonic mucosa of opossums contained uroguanylin and guanylin peptides, by purifying prouroguanylin and proguanylin from this tissue. Prouroguanylin and proguanylin coeluted from Sephadex G-75 gel- filtration columns with a similar molecular size between 6 and 12 kDa. Mass spectrometry indicated that proguanylin (≃8.7 kDa) had a 10% lower molecular mass than prouroguanylin (≃9.7 kDa). Isoelectric focusing separated prouroguanylin (pI ≃4.5) from proguanylin (pI ≃7.5). N-terminal sequence analysis of reverse phase-HPLC purified prohormones revealed 13 amino acids in opossum proguanylin that shared 77-85% identity with human and rat proguanylin, but only 23% identity with opossum prouroguanylin. The N- terminal 19 residues obtained for opossum prouroguanylin shared 32-42% identity with rat and human proguanylin. Prouroguanylin and proguanylin were both inactive and required pretreatment with proteases to elicit cyclic GMP responses in T84 cells. V8 protease treatment of proguanylin liberated a bioactive, 16-amino acid form of guanylin. Chymotrypsin treatment activated prouroguanylin, but inactivated the bioactive peptide domain within proguanylin. In summary, colonic mucosa contains the bioactive peptide and inactive prohormone forms of uroguanylin and guanylin. Thus, after proteolytic processing of prouroguanylin and proguanylin, bioactive uroguanylin and guanylin could both function to regulate guanylate cyclase activity by autocrine and/or paracrine actions on enterocytes. Also, these peptide hormones are implicated in an intestinal-renal axis for the endocrine regulation of salt and water homeostasis.

UR - http://www.scopus.com/inward/record.url?scp=0030025049&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030025049&partnerID=8YFLogxK

U2 - 10.1210/en.137.1.257

DO - 10.1210/en.137.1.257

M3 - Article

C2 - 8536621

AN - SCOPUS:0030025049

VL - 137

SP - 257

EP - 265

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 1

ER -