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Furin, PC1/3, and/or PC6A process rabbit, but not human, pro-lactase-phlorizin hydrolase to the 180-kDa intermediate.

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  • 1995-10-27
Published in:
  • The Journal of biological chemistry. - 1995
Amino Acid Sequence
Animals
Aspartic Acid Endopeptidases
Brefeldin A
Cloning, Molecular
Cyclopentanes
Enzyme Precursors
Furin
Humans
In Situ Hybridization
Intestine, Small
Lactase-Phlorizin Hydrolase
Mice
Molecular Sequence Data
Organ Culture Techniques
Polymerase Chain Reaction
Proprotein Convertase 1
Proprotein Convertase 5
Proprotein Convertases
Protein Processing, Post-Translational
RNA, Messenger
Rabbits
Rats
Recombinant Proteins
Sequence Analysis, DNA
Serine Endopeptidases
Species Specificity
Substrate Specificity
Subtilisins
Tissue Distribution
Transfection
English Small intestinal lactase-phlorizin hydrolase (LPH) is synthesized as a large precursor (prepro-LPH) of 1926 amino acids. In the endoplasmic reticulum, prepro-LPH is split by signal protease. The resulting pro-LPH is cut to mature LPH directly (human) or via a 180-kDa intermediate (rabbit), most likely in the trans-Golgi network or in a later compartment. Antibodies directed against different regions of rabbit pro-LPH locate the cleavage site resulting in the 180-kDa intermediate between amino acid residues 79 and 286. This stretch contains the two sequences -Arg-Cys-Tyr-Arg114 approximately -Arg-Ala-Ser-Arg191 approximately, which are potential cleavage sites for subtilisin-like proprotein convertases. These sites are not conserved in human pro-LPH. By coexpression in COS 7 cells of rabbit prepro-LPH and proprotein convertases (PC 1/3, PC2, PC6A, PC6B, furin), we show that furin, PC 1/3, and PC6A generate a processing intermediate that is immunologically indistinguishable from the one observed in vivo. Furin, PC 1/3, and PC6A are all expressed in the small intestine as shown by a polymerase chain reaction-based approach and, more specifically, in enterocytes, as shown by in situ hybridization. These results suggest that furin, PC 1/3, and/or PC6A are responsible for the in vivo processing of rabbit pro-LPH to the 180-kDa intermediate.
Language
  • English
Open access status
bronze
Identifiers
Persistent URL
https://sonar.ch/global/documents/191011
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