Multiple folding pathways for heterologously expressed human prion protein.


Abstract

Human PrP (residues 91-231) expressed in Escherichia coli can adopt several conformations in solution depending on pH, redox conditions and denaturant concentration. Oxidised PrP at neutral pH, with the disulphide bond intact, is a soluble monomer which contains 47% alpha-helix and corresponds to PrPC. Denaturation studies show that this structure has a relatively small, solvent-excluded core and unfolds to an unstructured state in a single, co-operative transition with a DeltaG for folding of -5.6 kcal mol-1. The unfolding behaviour is sensitive to pH and at 4.0 or below the molecule unfolds via a stable folding intermediate. This equilibrium intermediate has a reduced helical content and aggregates over several hours. When the disulphide bond is reduced the protein adopts different conformations depending upon pH. At neutral pH or above, the reduced protein has an alpha-helical fold, which is identical to that observed for the oxidised protein. At pH 4 or below, the conformation rearranges to a fold that contains a high proportion of beta-sheet structure. In the reduced state the alpha- and beta-forms are slowly inter-convertible whereas when oxidised the protein can only adopt an alpha-conformation in free solution. The data we present here shows that the human prion protein can exist in multiple conformations some of which are known to be capable of forming fibrils. The precise conformation that human PrP adopts and the pathways for unfolding are dependent upon solvent conditions. The conditions we examined are within the range that a protein may encounter in sub-cellular compartments and may have implications for the mechanism of conversion of PrPC to PrPSc in vivo. Since the conversion of PrPC to PrPSc is accompanied by a switch in secondary structure from alpha to beta, this system provides a useful model for studying major structural rearrangements in the prion protein. Study holds ProTherm entries: 15094, 15095, 15096, 15097, 15098 Extra Details: Folding to Unfolding prion; protein folding; thermodynamics; circular dichroism; recombinant expression; NMR

Submission Details

ID: NfUgQ3sN3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:45 p.m.

Version: 1

Publication Details
Jackson GS;Hill AF;Joseph C;Hosszu L;Power A;Waltho JP;Clarke AR;Collinge J,Biochim. Biophys. Acta (1999) Multiple folding pathways for heterologously expressed human prion protein. PMID:10209273
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1B10 1998-11-25T00:00:00+0000 0 SOLUTION NMR STRUCTURE OF RECOMBINANT SYRIAN HAMSTER PRION PROTEIN RPRP(90-231) , 25 STRUCTURES
2KKG 2009-06-19T00:00:00+0000 0 NMR structure of the octarepeat region of prion protein bound to pentosan polysulfate
2LH8 2011-08-05T00:00:00+0000 0 Syrian hamster prion protein with thiamine
3NVE 2010-07-08T00:00:00+0000 1.7 MMHFGN segment 138-143 from Syrian Hamster prion
4YXL 2015-03-23T00:00:00+0000 2.6 Crystal structure of Syrian hamster prion protein complexed with POM1 FAB
7LNA 2021-02-06T00:00:00+0000 3.1 Infectious mammalian prion fibril (263K scrapie)
1G04 2000-10-05T00:00:00+0000 0 SOLUTION STRUCTURE OF SYNTHETIC 26-MER PEPTIDE CONTAINING 145-169 SHEEP PRION PROTEIN SEGMENT AND C-TERMINAL CYSTEINE
1M25 2002-06-21T00:00:00+0000 0 STRUCTURE OF SYNTHETIC 26-MER PEPTIDE CONTAINING 145-169 SHEEP PRION PROTEIN SEGMENT AND C-TERMINAL CYSTEINE IN TFE SOLUTION
1S4T 2004-01-18T00:00:00+0000 0 Solution structure of synthetic 21mer peptide spanning region 135-155 (in human numbering) of sheep prion protein
1TPX 2004-06-16T00:00:00+0000 2.56 Ovine recombinant PrP(114-234), ARQ variant in complex with the Fab of the VRQ14 antibody

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
93.1 Major prion protein P04925 PRIO_MOUSE
91.7 Major prion protein P04273 PRIO_MESAU
92.9 Major prion protein P40249 PRIO_SAPAP
92.3 Major prion protein P40258 PRIO_SAISC
92.2 Major prion protein Q60506 PRIO_CRIGR
92.2 Major prion protein P13852 PRIO_RAT
90.4 Major prion protein P23907 PRIO_SHEEP
90.4 Major prion protein Q5XVM4 PRIO_RUPRU
90.4 Major prion protein Q7JK02 PRIO_OVIMU
90.4 Major prion protein Q7JIY2 PRIO_OVIMO
90.4 Major prion protein Q7JIH3 PRIO_OVICA
90.4 Major prion protein P52113 PRIO_CAPHI
93.4 Major prion protein P40246 PRIO_ATEGE
90.9 Major prion protein Q95M08 PRIO_BUDTA
93.6 Major prion protein Q9Z0T3 PRIO_SIGHI
95.3 Major prion protein P51446 PRIO_ATEPA
95.3 Major prion protein P40247 PRIO_CALJA
96.6 Major prion protein P40257 PRIO_TRAFR
91.3 Major prion protein Q95270 PRIO_THEGE
95.4 Major prion protein P40245 PRIO_AOTTR
93.1 Major prion protein P67991 PRIO_MACSY
93.1 Major prion protein P67990 PRIO_LOPAT
92.9 Major prion protein P67988 PRIO_CHLAE
92.9 Major prion protein P67989 PRIO_CERDI
95.9 Major prion protein Q95176 PRIO_CERAT
95.9 Major prion protein P40255 PRIO_MANSP
95.9 Major prion protein P40248 PRIO_PLEMO
96.4 Major prion protein P67996 PRIO_PAPHA
96.4 Major prion protein P67995 PRIO_MACNE
96.4 Major prion protein P67997 PRIO_MACMU
96.4 Major prion protein P67994 PRIO_MACFU
96.4 Major prion protein P67992 PRIO_MACFA
96.4 Major prion protein P67993 PRIO_MACAR
95.9 Major prion protein Q95174 PRIO_ERYPA
96.3 Major prion protein P61762 PRIO_CERNE
96.3 Major prion protein P61761 PRIO_CERMO
97.2 Major prion protein P40251 PRIO_COLGU
98.1 Major prion protein P40256 PRIO_PONPY
99.2 Major prion protein P61767 PRIO_SYMSY
99.2 Major prion protein P61768 PRIO_PANTR
99.2 Major prion protein P61766 PRIO_HYLLA
99.6 Major prion protein P40252 PRIO_GORGO
100.0 Major prion protein P04156 PRIO_HUMAN