Analysis of the human guanylin gene and the processing and cellular localization of the peptide.
The complete cell biological analysis of human guanylin, a recently discovered regulatory peptide, is offered in this investigation: (i) the nucleotide sequence of the gene, (ii) the isolation and characterization of its circulating molecular form, and (iii) its localization in enterochromaffin cells of the gut. As determined by molecular cloning, DNA sequencing, and comparison with the known cDNA sequence, the approximately 2.6-kbp large gene consists of three exons interrupted by two introns. The putative promoter region contains a TTTAAAA sequence motif and several potential binding sites for transcription factors such as AP-1, AP-2, Sp 1, and glucocorticoid receptors. The isolated hormonal form of guanylin is a 94-amino acid peptide with a molecular mass of 10.3 kDa. Western blot analysis of RP-HPLC fractions from blood plasma confirms this molecular form. Thus, guanylin is synthesized by gut enterochromaffin cells as a prohormone of 115 amino acids and is processed to the molecular form of 94 amino acids circulating in the blood.
Hill
O
O
Lower Saxony Institute for Peptide Research, Hannover, Germany.
Kuhn
M
M
Zucht
H D
HD
Cetin
Y
Y
Kulaksiz
H
H
Adermann
K
K
Klock
G
G
Rechkemmer
G
G
Forssmann
W G
WG
Mägert
H J
HJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002034:Bumetanide / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003106:Colon; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005768:Gastrointestinal Hormones; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000502:physiology; D007438:Introns; D008969:Molecular Sequence Data; D008970:Molecular Weight; D045265:Natriuretic Peptides; D010452:Peptide Biosynthesis; D010455:Peptides / Q000235:genetics* / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D016329:Sp1 Transcription Factor / Q000378:metabolism; D016385:TATA Box; D018808:Transcription Factor AP-1 / Q000378:metabolism; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000378:metabolism
D017931:DNA Primers; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D005768:Gastrointestinal Hormones; D045265:Natriuretic Peptides; D010455:Peptides; D011965:Receptors, Glucocorticoid; D011994:Recombinant Proteins; D016329:Sp1 Transcription Factor; D018808:Transcription Factor AP-1; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D002034:Bumetanide; C072685:guanylin