Enhanced stability of human prion proteins with two disulfide bridges.

Abstract

We compare the folding equilibrium of the globular domain of the human prion protein with two variants of this domain, for which an additional disulfide bond was introduced into the location where it is found in the naturally occurring doppel protein. We find that the unfolding transition midpoint of the variants is shifted toward higher denaturant concentration, indicating that the engineered disulfide bond significantly stabilizes the global protein structure. Our results further reveal that the two-disulfide variant proteins, while possessing the same global fold as the wild-type, display marked differences in their folding pathway-in particular, the absence of a characteristic alpha-helix to beta-sheet transition, which is a fundamental feature associated with misfolding of proteins into amyloid fibrils, especially in the context of prion diseases. These surprising characteristics of disulfide mutant prion proteins have important implications for the understanding of the generic aberrant processes leading to amyloid fibril formation and protein aggregation, as well as providing insight into possible therapeutic strategies. Study holds ProTherm entries: 22279, 22280, 22281, 22282 Extra Details: disulfide bond; folding pathway; amyloid fibrils; transition

Submission Details

ID: gYXY3gKU3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:53 p.m.

Version: 1

Publication Details
Knowles TP;Zahn R,Biophys. J. (2006) Enhanced stability of human prion proteins with two disulfide bridges. PMID:16751235
Additional Information

Study Summary

Number of data points 5
Proteins Major prion protein ; Major prion protein
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Tm ; Experimental Assay: Cm pH:5.0, buffers:sodium acetate: 20mM ; Experimental Assay: Cm buffers:sodium phosphate: 20mM, pH:7.0 ; Experimental Assay: dG_H2O
Libraries Mutations for sequence GSKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHSQWNKPSKPKTNMKHMAGAAAAGAVVGGLGGYMLGSAMSRPIIHFGSDYEDRYYRENMHRYPNQVYYRPMDEYSNQNNFVHDCVNITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYERESQAYYQRGS

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 UniProtKB Entries

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