Molecular cloning and cellular localization of a furin-like prohormone convertase from the atrial gland of Aplysia.
Prohormone convertases (PCs) are Ca(2+)-dependent subtilisin-related endoproteases that have been implicated in the post-translational processing of prohormones and other proproteins. Furin is an ubiquitously expressed PC that has been shown to hydrolyze a wide variety of precursor proteins in secretory pathways. We have screened an Aplysia atrial gland cDNA library using a furin probe prepared by polymerase chain reaction (PCR) and have isolated an Aplysia furin-related 6.7-kb cDNA partial clone that was truncated on the 5' end. The remaining 5' atrial gland furin nucleotide sequence was obtained by two stages of reverse transcription PCR. The final composite nucleotide sequence of the atrial gland furin cDNA was 7,837 bp in length. This sequence encoded a putative preproendoprotease (Afurin2) of 824 amino acid residues that was related to other eukaryotic furins, and showed a high sequence identity with a recently reported Aplysia nervous system furin-like sequence. In situ hybridization demonstrated extensive expression of Afurin2 in atrial gland secretory cells. The cDNA clone contained a relatively long 3' untranslated region of 5,230 nucleotides that included a microsatellite repeat region (TG)n. The characterized Aplysia Afurin2 is a candidate PC that may play an important role in the processing of egg-laying hormone (ELH)-related precursors in the secretory cells of the atrial gland. In addition, comparative structural studies of Afurin2, together with previously reported localization studies, argue for the occurrence of a furin-like convertase within secretory granules.
Nagle
G T
GT
Department of Anatomy and Neurosciences, University of Texas Medical Branch, Galveston 77555, USA.
Garcia
A T
AT
Gorham
E L
EL
Knock
S L
SL
van Heumen
W R
WR
Spijker
S
S
Smit
A B
AB
Geraerts
W P
WP
Kurosky
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia / Q000201:enzymology*; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D045683:Furin; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D013381:Subtilisins / Q000235:genetics* / Q000378:metabolism
D013381:Subtilisins; C093644:afurin 2; D045683:Furin