Functional analysis of an ADAMTS10 signal peptide mutation in Weill-Marchesani syndrome demonstrates a long-range effect on secretion of the full-length enzyme.

Abstract

We report the identification and functional analysis of the first missense ADAMTS10 mutation (c.73G>A; p.Ala25Thr) causing recessive Weill-Marchesani syndrome (WMS). The Ala25 residue affected by the missense mutation is at the -1 position relative to the ADAMTS10 signal peptidase cleavage site. p.Ala25Thr substituted full-length ADAMTS10 showed consistent and significantly diminished secretion in both HEK293F and Cos-1 cells. However, a C-terminally truncated construct lacking the ancillary domain and containing only the signal peptide, the propeptide and the catalytic domain (p.Ala25Thr Pro-Cat) was efficiently secreted in both HEK293F cells and Cos-1 cells. Edman degradation of purified p.Ala25Thr Pro-Cat and p.Ala25Thr substituted full-length ADAMTS10 from HEK293F cells demonstrated correct signal peptide processing. Thus, the p.Ala25Thr substitution hinders secretion of full-length ADAMTS10, but not Pro-Cat from cells, yet permits signal peptide removal. We infer that folding of the complex C-terminal ancillary domain is the rate-limiting step in biosynthesis of ADAMTS10, and that it (but not Pro-Cat) is sensitive to subtle changes in efficiency of signal peptide cleavage. These observations represent an unprecedented effect of a signal peptide mutation and support a model in which the initial cotranslational processing events during protein biosynthesis can have long-range effects on protein folding and secretion.

• Authors: Wendy E Kutz, Lauren W Wang, Nathalie Dagoneau, Kazimir J Odrcic, Valerie Cormier-Daire, Elias I Traboulsi, Suneel S Apte
• Journal: Human mutation (Hum Mutat) Vol. 29 Issue 12 Pg. 1425-34 (Dec 2008) ISSN: 1098-1004 [Electronic] United States
• PMID: 18567016 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Protein Sorting Signals
• ADAM Proteins
• ADAMTS Proteins
• ADAMTS10 protein, human

Topics

• ADAM Proteins (chemistry, genetics)
• ADAMTS Proteins
• Aged, 80 and over
• Animals
• COS Cells
• Cell Line
• Chlorocebus aethiops
• DNA Mutational Analysis
• Growth Disorders (genetics)
• Humans
• Mutation
• Protein Sorting Signals
• Protein Structure, Tertiary
• Syndrome