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
2LEJ 2011-08-17 human prion protein mutant HuPrP(90-231, M129, V210I)
2RMW 2007-12-18 Solution structure of synthetic 26-mer peptide containing 142-166 sheep prion protein segment and C-terminal cysteine with R156A mutation
2KFO 2009-06-16 Mouse Prion Protein (121-231) with Mutation V166A
1XYU 2005-01-04 Solution structure of the sheep prion protein with polymorphism H168
1Y2S 2004-12-28 Ovine Prion Protein Variant R168
2KU5 2010-06-09 Mouse Prion Protein (121-231) with mutation D167S
1Y16 2005-01-25 mouse prion protein with mutations S170N and N174T
2K1D 2009-03-03 NMR Studies of a Pathogenic Mutant (D178N) of the Human Prion Protein
2KKG 2009-11-10 NMR structure of the octarepeat region of prion protein bound to pentosan polysulfate
1QM2 1999-12-16 Human prion protein fragment 121-230
1HJM 2003-07-03 HUMAN PRION PROTEIN AT PH 7.0
1E1J 2000-07-20 Human prion protein variant M166V
5YJ4 2018-04-11 structure for the protective mutant G127V of Human prion protein
2IV6 2006-11-28 hPrP-173-195-D178N solution structure
5YJ5 2018-04-11 structure for wildtype Human prion protein (M129)
2L39 2011-08-10 Mouse prion protein fragment 121-231 AT 37 C
1OEI 2004-05-06 Human prion protein 61-84
1H0L 2003-01-30 HUMAN PRION PROTEIN 121-230 M166C/E221C
1E1S 2000-07-21 Human prion protein variant S170N
1E1W 2000-07-20 Human prion protein variant R220K
2KFM 2009-06-16 Mouse Prion Protein (121-231) with Mutations Y225A and Y226A
2LBG 2011-12-07 Structure of the CHR of the Prion protein in DPC Micelles
1B10 1998-12-02 SOLUTION NMR STRUCTURE OF RECOMBINANT SYRIAN HAMSTER PRION PROTEIN RPRP(90-231) , 25 STRUCTURES
1G04 2002-01-23 SOLUTION STRUCTURE OF SYNTHETIC 26-MER PEPTIDE CONTAINING 145-169 SHEEP PRION PROTEIN SEGMENT AND C-TERMINAL CYSTEINE
1M25 2002-07-17 STRUCTURE OF SYNTHETIC 26-MER PEPTIDE CONTAINING 145-169 SHEEP PRION PROTEIN SEGMENT AND C-TERMINAL CYSTEINE IN TFE SOLUTION
1FO7 2000-09-21 HUMAN PRION PROTEIN MUTANT E200K FRAGMENT 90-231
2MV8 2015-10-07 Solution structure of Ovis Aries PrP with mutation delta190-197
2L1K 2011-08-10 Mouse prion protein (121-231) containing the substitutions Y169A, Y225A, and Y226A
2L1E 2011-08-10 Mouse prion protein (121-231) containing the substitution F175A
2L1H 2011-08-10 Mouse prion protein fragment 121-231 at 20 C
1Y15 2004-12-28 Mouse Prion Protein with mutation N174T
1S4T 2004-01-27 Solution structure of synthetic 21mer peptide spanning region 135-155 (in human numbering) of sheep prion protein
2LFT 2012-06-27 Human prion protein with E219K protective polymorphism
2LV1 2012-09-19 Solution-state NMR structure of prion protein mutant V210I at neutral pH
2MV9 2015-10-07 Solution structure of Ovis Aries PrP with mutation delta193-196
2IV5 2006-11-28 hPrP-173-195 solution structure
1QM1 1999-12-16 Human prion protein fragment 90-230
2L40 2011-08-10 Mouse prion protein (121-231) containing the substitution Y169A
2M8T 2013-09-11 Solution NMR structure of the V209M variant of the human prion protein (residues 90-231)
1E1G 2000-07-20 Human prion protein variant M166V
2LSB 2012-06-27 Solution-state NMR structure of the human prion protein
1AG2 1997-10-08 PRION PROTEIN DOMAIN PRP(121-231) FROM MOUSE, NMR, 2 MINIMIZED AVERAGE STRUCTURE
1QM3 1999-12-16 Human prion protein fragment 121-230
1FKC 2000-09-21 HUMAN PRION PROTEIN (MUTANT E200K) FRAGMENT 90-231
1E1P 2000-07-20 Human prion protein variant S170N
1HJN 2003-07-03 HUMAN PRION PROTEIN AT PH 7.0
5L6R 2016-10-05 PrP226* - Solution-state NMR structure of truncated human prion protein
2IV4 2006-11-28 hPrP180-195 structure
2K5O 2008-12-09 Mouse Prion Protein (121-231) with Mutation S170N
1E1U 2000-07-20 Human prion protein variant R220K
1QM0 1999-12-16 Human prion protein fragment 90-230
2LH8 2011-09-14 Syrian hamster prion protein with thiamine
1QLX 1999-12-16 Human prion protein
2N53 2016-05-25 Solution structure of ovis aries prp
2RMV 2007-12-18 Solution structure of synthetic 26-mer peptide containing 142-166 sheep prion protein segment and C-terminal cysteine with Y155A mutation
1QLZ 1999-12-16 Human prion protein
1OEH 2004-04-23 Human prion protein 61-68
1XYX 2004-11-28 mouse prion protein fragment 121-231
2KU6 2010-06-09 Mouse Prion Protein (121-231) with mutations D167S and N173K
2L1D 2011-08-10 Mouse prion protein (121-231) containing the substitution Y169G
2KUN 2010-08-25 Three dimensional structure of HuPrP(90-231 M129 Q212P)
2KTM 2010-04-07 Solution NMR structure of H2H3 domain of ovine prion protein (residues 167-234)
3MD4 2011-05-25 1.15 Prion peptide
4E1H 2013-03-06 1.4 Fragment of human prion protein
3MD5 2011-05-25 1.4 Prion peptide
3NVG 2011-03-02 1.48 MIHFGN segment 137-142 from mouse prion
4N9O 2014-01-22 1.5 Probing the N-terminal beta-sheet conversion in the crystal structure of the human prion protein bound to a Nanobody
4KML 2014-02-19 1.5 Probing the N-terminal beta-sheet conversion in the crystal structure of the full-length human prion protein bound to a Nanobody
3NHC 2010-08-04 1.57 GYMLGS segment 127-132 from human prion with M129
3NVH 2011-03-02 1.61 MIHFGND segment 137-143 from mouse prion
3NVE 2011-03-02 1.7 MMHFGN segment 138-143 from Syrian Hamster prion
3HAK 2010-01-12 1.8 Human prion protein variant V129
3HEQ 2010-01-12 1.8 Human prion protein variant D178N with M129
3NVF 2011-03-02 1.8 IIHFGS segment 138-143 from human prion
6AQ7 2018-04-04 1.83 Structure of POM6 FAB fragment complexed with mouse PrPc
3HER 2010-01-12 1.85 Human prion protein variant F198S with V129
4H88 2013-07-31 1.9 Structure of POM1 FAB fragment complexed with mouse PrPc Fragment 120-230
3NHD 2010-08-04 1.92 GYVLGS segment 127-132 from human prion with V129
4MA7 2014-01-22 1.97 Crystal structure of mouse prion protein complexed with Promazine
3HES 2010-01-12 2.0 Human prion protein variant F198S with M129
3HJX 2010-01-12 2.0 Human prion protein variant D178N with V129
1I4M 2002-02-27 2.0 Crystal structure of the human prion protein reveals a mechanism for oligomerization
4E1I 2013-03-06 2.03 Fragment of human prion protein
1UW3 2004-03-25 2.05 The crystal structure of the globular domain of sheep prion protein
4MA8 2014-01-22 2.2 Crystal structure of mouse prion protein complexed with Chlorpromazine
3HAF 2010-01-12 2.26 Human prion protein variant V129 domain swapped dimer
4J8R 2013-05-22 2.3 Structure of an octapeptide repeat of the prion protein bound to the POM2 Fab antibody fragment
6DU9 2019-07-03 2.33 Crystal Structure of Human Prion Protein 90-231
4DGI 2012-10-31 2.4 Structure of POM1 FAB fragment complexed with human PrPc Fragment 120-230
1TQB 2004-07-06 2.55 Ovine recombinant PrP(114-234), VRQ variant in complex with the Fab of the VRQ14 antibody
1TPX 2004-07-06 2.56 Ovine recombinant PrP(114-234), ARQ variant in complex with the Fab of the VRQ14 antibody
4YXL 2015-09-23 2.6 Crystal structure of Syrian hamster prion protein complexed with POM1 FAB
1TQC 2004-07-06 2.8 Ovine recombinant PrP(114-234), ARR variant in complex with the VRQ14 Fab fragment (IgG2a)
2W9E 2009-02-03 2.9 Structure of ICSM 18 (anti-Prp therapeutic antibody) Fab fragment complexed with human Prp fragment 119-231
3HJ5 2010-01-12 3.1 Human prion protein variant V129 domain swapped dimer

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