Stability and global fold of the mouse prohormone convertase 1 pro-domain

Biochemistry
Michael A TangreaJohn Orban

Abstract

We have purified the mouse prohormone convertase 1 (PC1) pro-domain expressed in Escherichia coli cells and demonstrated, using a number of biophysical methods, that this domain is an independent folding unit with a T(m) of 39 degrees C at a protein concentration of 20 microM and pH 7.0. This differs significantly from similar pro-domains in bacteria and human furin, which are unfolded at 25 degrees C and require the catalytic domain in order to be structured [Bryan et al. (1995) Biochemistry 34, 10310-10318; Bhattacharjya et al. (2000) J. Biomol. NMR 16, 275-276]. Using heteronuclear NMR spectroscopy, we have determined the backbone (1)H, (13)C, and (15)N assignments for the pro-domain of PC1. On the basis of (1)H/(13)C chemical shift indices, NOE analysis, and hydrogen exchange measurements, the pro-domain is shown to consist of a four-stranded beta-sheet and two alpha-helices. The results presented here show that both the bacterial pro-domain in complex with subtilisin and the uncomplexed mouse PC1 pro-domain have very similar overall folds despite a lack of sequence homology. The structural data help to explain the location of the secondary processing sites in the pro-domains of the PC family, and a consensus sequence for b...Continue Reading

References

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Citations

Jul 4, 2002·Journal of Molecular Biology·Michael A TangreaJohn Orban
Mar 23, 2002·Protein Science : a Publication of the Protein Society·Vladimir N Uversky
May 5, 2007·Biopolymers·Surajit BhattacharjyaFeng Ni
Jun 22, 2005·Expert Review of Proteomics·Michael A TangreaRodrigo F Chuaqui
Apr 8, 2006·The Journal of Biological Chemistry·Sylvain F FeliciangeliGary Thomas
Nov 28, 2001·The Journal of Biological Chemistry·Martin FugèreRobert Day
Oct 19, 2005·The Biochemical Journal·Sergey A ShiryaevAlex Y Strongin
May 24, 2003·The Journal of Biological Chemistry·Letitia JeanMichael J Blackman
May 9, 2013·The Journal of Biological Chemistry·Danielle M WilliamsonUjwal Shinde
Dec 12, 2002·Chemical Reviews·Philip N Bryan

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