Mutations D187N and D187Y in domain 2 of the actin-regulating protein gelsolin cause familial amyloidosis-Finnish type (FAF). We have constructed and expressed a recombinant version of gelsolin domain 2 that is sufficiently stable for kinetic and equilibrium measurements. The wild-type domain and the two amyloidogenic mutants fold via simple two-state kinetics without the accumulation of an intermediate. Unfolding kinetics exhibits significant curvature with increasing urea concentration, indicating that the transition state for unfolding becomes more native-like under increasingly denaturing conditions in accordance with the Hammond postulate. Mutations D187N and D187Y destabilize gelsolin domain 2 by 1.22 and 2.16 kcal. mol(-1) (1 kcal = 4.18 kJ) respectively. The mutations do not prevent disulfide bond formation despite their direct contiguity with a cysteine residue involved in disulfide linkage. The destabilization conferred on gelsolin domain 2 by the FAF mutations is sufficient to predict that an appreciable fraction is unfolded and, therefore, extremely susceptible to proteolysis at body temperature. Study holds ProTherm entries: 6490, 6491, 6492, 6493 Extra Details: protein; stability; denaturation; Hammond; two-state
ID: ezWdA95J
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:32 p.m.
Version: 1
Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
---|
Percent Identity | Matching Chains | Protein | Accession | Entry Name |
---|---|---|---|---|
99.5 | A | Major actin | P07830 | ACT1_DICDI |
99.2 | A | Major actin | Q54GX7 | ACT10_DICDI |
98.9 | A | Major actin | P02576 | ACTA_PHYPO |
98.4 | A | Major actin | Q553U6 | ACT22_DICDI |
97.6 | A | Major actin | P02578 | ACT1_ACACA |
96.5 | A | Major actin | P07829 | ACT3_DICDI |
95.7 | A | Major actin | O93400 | ACTB_XENLA |
95.7 | A | Major actin | Q6NVA9 | ACTB_XENTR |
95.7 | A | Major actin | P10987 | ACT1_DROME |
95.7 | A | Major actin | P84184 | ACT3B_HELAM |
95.7 | A | Major actin | P84183 | ACT4_BOMMO |
95.7 | A | Major actin | P84185 | ACT5C_ANOGA |
95.2 | A | Major actin | P10984 | ACT2_CAEEL |
95.2 | A | Major actin | P53471 | ACT2_SCHMA |
95.7 | A | Major actin | P53505 | ACT5_XENLA |
95.4 | A | Major actin | P60712 | ACTB_BOVIN |
95.4 | A | Major actin | O18840 | ACTB_CANLF |
95.4 | A | Major actin | Q71FK5 | ACTB_CAVPO |
95.4 | A | Major actin | P60706 | ACTB_CHICK |
95.4 | A | Major actin | Q76N69 | ACTB_CHLAE |
95.4 | A | Major actin | P60708 | ACTB_HORSE |
95.4 | A | Major actin | P60709 | ACTB_HUMAN |
95.4 | A | Major actin | Q4R561 | ACTB_MACFA |
95.4 | A | Major actin | Q711N9 | ACTB_MESAU |
95.4 | A | Major actin | P60710 | ACTB_MOUSE |
95.4 | A | Major actin | Q5R1X3 | ACTB_PANTR |
95.4 | A | Major actin | Q6QAQ1 | ACTB_PIG |
95.4 | A | Major actin | Q5R6G0 | ACTB_PONAB |
95.4 | A | Major actin | P60711 | ACTB_RAT |
95.4 | A | Major actin | P60713 | ACTB_SHEEP |
95.4 | A | Major actin | Q4L0Y2 | ACTB_SPECI |
95.4 | A | Major actin | P60707 | ACTB_TRIVU |
95.7 | A | Major actin | P63258 | ACTG_BOVIN |
95.7 | A | Major actin | Q5ZMQ2 | ACTG_CHICK |
95.7 | A | Major actin | P63261 | ACTG_HUMAN |
95.7 | A | Major actin | P63260 | ACTG_MOUSE |
95.7 | A | Major actin | Q5JAK2 | ACTG_PELLE |
95.7 | A | Major actin | P63259 | ACTG_RAT |
95.7 | A | Major actin | P63257 | ACTG_TRIVU |
95.7 | A | Major actin | A2BDB0 | ACTG_XENLA |
95.7 | A | Major actin | Q25010 | ACT3A_HELAM |
95.2 | A | Major actin | Q93129 | ACTC_BRABE |
95.4 | A | Major actin | Q6P378 | ACTG_XENTR |
95.2 | A | Major actin | P41340 | ACT3_LIMPO |
95.4 | A | Major actin | P18603 | ACT4_ARTSX |
96.0 | A | Major actin | P12716 | ACTC_PISOC |
94.9 | A | Major actin | P30162 | ACT1_ONCVO |
95.2 | A | Major actin | P02572 | ACT2_DROME |
95.2 | A | Major actin | P53478 | ACT5_CHICK |
95.2 | A | Major actin | P48975 | ACTB_CRIGR |
95.2 | A | Major actin | P30163 | ACT2_ONCVO |
95.2 | A | Major actin | P68142 | ACTB1_TAKRU |
95.2 | A | Major actin | P53486 | ACTB3_TAKRU |
95.2 | A | Major actin | P68143 | ACTB_OREMO |
95.2 | A | Major actin | Q91ZK5 | ACTB_SIGHI |
94.9 | A | Major actin | Q93131 | ACTC_BRAFL |
95.2 | A | Major actin | Q8JJB8 | ACTG_TRISC |
94.6 | A | Major actin | P10986 | ACT4_CAEEL |
94.9 | A | Major actin | P79818 | ACTB_ORYLA |
95.4 | A | Major actin | P15475 | ACTB_XENBO |
95.4 | A | Major actin | P04829 | ACT3_BOMMO |
94.4 | A | Major actin | P0DM41 | ACT1_CAEEL |
94.4 | A | Major actin | P0DM42 | ACT3_CAEEL |
95.4 | A | Major actin | P53506 | ACT8_XENLA |
94.9 | A | Major actin | O42161 | ACTB_SALSA |
94.9 | A | Major actin | Q7ZVI7 | ACTB1_DANRE |
95.2 | A | Major actin | P29751 | ACTB_RABIT |
95.2 | A | Major actin | Q26065 | ACT_PLAMG |
94.4 | A | Major actin | O17320 | ACT_CRAGI |
94.6 | A | Major actin | Q7ZVF9 | ACTB2_DANRE |
94.6 | A | Major actin | P83751 | ACTB_CTEID |
94.6 | A | Major actin | P83750 | ACTB_CYPCA |
94.9 | A | Major actin | Q964E1 | ACTC_BIOOB |
94.9 | A | Major actin | Q964E2 | ACTC_BIOPF |
94.4 | A | Major actin | O18499 | ACT1_SACKO |
94.9 | A | Major actin | P53470 | ACT1_SCHMA |
94.6 | A | Major actin | Q964E3 | ACTC_BIOAL |
95.2 | A | Major actin | P63256 | ACTG_ANSAN |
94.4 | A | Major actin | P53485 | ACTB2_TAKRU |
94.9 | A | Major actin | P53461 | ACTC_HALRO |
94.6 | A | Major actin | Q0PGG4 | ACTB_BOSMU |
94.6 | A | Major actin | P92179 | ACTC_BIOGL |
94.4 | A | Major actin | Q964E0 | ACTC_BIOTE |
94.4 | A | Major actin | Q964D9 | ACTC_PLATR |
94.9 | A | Major actin | P90689 | ACT_BRUMA |
94.9 | A | Major actin | P84336 | ACTB_CAMDR |
94.4 | A | Major actin | P17126 | ACT_HYDVU |
94.6 | A | Major actin | P41339 | ACTA_LIMPO |
94.1 | A | Major actin | P49871 | ACT_MANSE |
93.8 | A | Major actin | P07836 | ACT1_BOMMO |
93.8 | A | Major actin | P68556 | ACT1_DIPDE |
94.9 | A | Major actin | P41112 | ACT1_PODCA |
93.8 | A | Major actin | P68555 | ACT_TAESO |
93.6 | A | Major actin | P45886 | ACT3_BACDO |
94.1 | A | Major actin | P41113 | ACT3_PODCA |
93.6 | A | Major actin | P45885 | ACT2_BACDO |
93.6 | A | Major actin | P92182 | ACT1_LUMTE |
93.6 | A | Major actin | O17503 | ACTC_BRALA |
93.6 | A | Major actin | P53456 | ACT2_DIPDE |
93.3 | A | Major actin | P53501 | ACT3_DROME |
94.1 | A | Major actin | O18500 | ACT2_SACKO |
93.3 | A | Major actin | P10981 | ACT5_DROME |
94.1 | A | Major actin | P41341 | ACTY_LIMPO |
94.9 | A | Major actin | P69002 | ACT1_HELER |
94.9 | A | Major actin | P69003 | ACT1_HELTB |
94.3 | A | Major actin | P53458 | ACT5_DIPDE |
93.3 | A | Major actin | P18601 | ACT2_ARTSX |
94.1 | A | Major actin | P92176 | ACT2_LUMTE |
93.3 | A | Major actin | P53464 | ACTM_HELTB |
93.0 | A | Major actin | O16808 | ACT_MAYDE |
93.8 | A | Major actin | P83969 | ACT1_BACDO |
93.8 | A | Major actin | P83967 | ACT6_DROME |
93.8 | A | Major actin | P83968 | ACT6_DROSI |
94.6 | A | Major actin | P53472 | ACTA_STRPU |
94.1 | A | Major actin | P17304 | ACTM_APLCA |
92.8 | A | Major actin | P18600 | ACT1_ARTSX |
93.3 | A | Major actin | P53463 | ACTM_HELER |
93.3 | A | Major actin | P07837 | ACT2_BOMMO |
93.3 | A | Major actin | P49128 | ACT1_AEDAE |
93.0 | A | Major actin | P53457 | ACT3_DIPDE |
93.3 | A | Major actin | P12717 | ACTM_PISOC |
93.3 | A | Major actin | P91754 | ACT_LUMRU |
94.6 | A | Major actin | P69004 | ACT2_MESFR |
94.6 | A | Major actin | P69005 | ACTD_STRPU |
91.7 | A | Major actin | P53496 | ACT11_ARATH |
94.6 | A | Major actin | P10990 | ACT1_MESFR |
91.5 | A | Major actin | A2XLF2 | ACT1_ORYSI |
91.5 | A | Major actin | Q10DV7 | ACT1_ORYSJ |
94.4 | A | Major actin | P53473 | ACTB_STRPU |
92.3 | A | Major actin | O65315 | ACT_COLSC |
93.6 | A | Major actin | P53465 | ACT1_LYTPI |
92.8 | A | Major actin | P45887 | ACT5_BACDO |
92.5 | A | Major actin | P62739 | ACTA_BOVIN |
92.5 | A | Major actin | P62736 | ACTA_HUMAN |
92.5 | A | Major actin | P62737 | ACTA_MOUSE |
92.5 | A | Major actin | P62740 | ACTA_RABIT |
92.5 | A | Major actin | P62738 | ACTA_RAT |
90.9 | A | Major actin | Q05214 | ACT1_TOBAC |
92.2 | A | Major actin | Q25472 | ACT2_MOLOC |
90.9 | A | Major actin | P30167 | ACT3_SOLTU |
92.5 | A | Major actin | P02574 | ACT4_DROME |
92.2 | A | Major actin | P08023 | ACTA_CHICK |
92.0 | A | Major actin | Q3ZC07 | ACTC_BOVIN |
92.0 | A | Major actin | P68034 | ACTC_CHICK |
92.0 | A | Major actin | P68032 | ACTC_HUMAN |
92.0 | A | Major actin | P68033 | ACTC_MOUSE |
92.0 | A | Major actin | P68035 | ACTC_RAT |
92.0 | A | Major actin | Q6P640 | ACTC_XENTR |
92.2 | A | Major actin | Q5E9B5 | ACTH_BOVIN |
92.2 | A | Major actin | P63270 | ACTH_CHICK |
92.2 | A | Major actin | P63267 | ACTH_HUMAN |
92.2 | A | Major actin | P63268 | ACTH_MOUSE |
92.2 | A | Major actin | P63269 | ACTH_RAT |
90.9 | A | Major actin | O81221 | ACT_GOSHI |
91.7 | A | Major actin | P20399 | ACT2_XENTR |
91.2 | A | Major actin | A2XNS1 | ACT3_ORYSI |
91.2 | A | Major actin | Q10AZ4 | ACT3_ORYSJ |
93.6 | A | Major actin | Q07903 | ACTC_STRPU |
92.0 | A | Major actin | P04751 | ACTC_XENLA |
90.7 | A | Major actin | P30171 | ACT11_SOLTU |
92.8 | A | Major actin | P53466 | ACT2_LYTPI |
92.0 | A | Major actin | P27130 | ACT2_HALRO |
91.7 | A | Major actin | P04752 | ACT3_XENLA |
91.7 | A | Major actin | Q6P8G3 | ACT3_XENTR |
91.7 | A | Major actin | P53459 | ACT6_DIPDE |
92.8 | A | Major actin | Q00215 | ACTC_STYPL |
92.2 | A | Major actin | O15998 | ACTM_CIOSA |
90.4 | A | Major actin | P0CJ46 | ACT1_ARATH |
90.4 | A | Major actin | P0CJ47 | ACT3_ARATH |
91.2 | A | Major actin | P53492 | ACT7_ARATH |
91.5 | A | Major actin | P53480 | ACTC_TAKRU |
92.8 | A | Major actin | P18499 | ACTF_STRPU |
91.2 | A | Major actin | P49055 | ACTS_CARAU |
93.0 | A | Major actin | P53474 | ACTE_STRPU |
91.2 | A | Major actin | P68138 | ACTS_BOVIN |
91.2 | A | Major actin | P68139 | ACTS_CHICK |
91.2 | A | Major actin | P68133 | ACTS_HUMAN |
91.2 | A | Major actin | P68134 | ACTS_MOUSE |
91.2 | A | Major actin | Q98972 | ACTS_ORYLA |
91.2 | A | Major actin | P68137 | ACTS_PIG |
91.2 | A | Major actin | Q5R9Q5 | ACTS_PONAB |
91.2 | A | Major actin | P68135 | ACTS_RABIT |
91.2 | A | Major actin | P68136 | ACTS_RAT |
91.2 | A | Major actin | P10995 | ACT2_XENLA |
91.8 | A | Major actin | P53475 | ACTN_STYCL |
91.2 | A | Major actin | P53482 | ACTSB_TAKRU |
90.9 | A | Major actin | P68140 | ACTSA_TAKRU |
90.9 | A | Major actin | P68264 | ACTS_OREMO |
90.6 | A | Major actin | P26197 | ACT2_ABSGL |
90.9 | A | Major actin | Q90X97 | ACTS_ATRMM |
90.7 | A | Major actin | P30173 | ACT13_SOLTU |
90.7 | A | Major actin | P53479 | ACTS_CYPCA |
91.2 | A | Major actin | P53460 | ACT1_HALRO |
90.1 | A | Major actin | P53494 | ACT4_ARATH |
91.5 | A | Major actin | P26198 | ACTM_STYCL |
91.4 | A | Major actin | P53689 | ACT_PHARH |
90.7 | A | Major actin | P53498 | ACT_CHLRE |
91.4 | A | Major actin | P20904 | ACT_VOLCA |
90.1 | A | Major actin | P53504 | ACT1_SORBI |
91.2 | A | Major actin | Q9Y707 | ACT2_SUIBO |
90.6 | A | Major actin | P50138 | ACT_PUCGR |
95.6 | A | Major actin | P84856 | ACTB_CHLPG |
97.2 | A | Major actin | Q55EU6 | ACT23_DICDI |
90.1 | A | Major actin | P53467 | ACTM_MOLOC |
90.3 | A | Major actin | Q9Y702 | ACT1_SCHCO |
90.4 | A | Major actin | P30164 | ACT1_PEA |
90.3 | A | Major actin | Q00214 | ACTM_STYPL |
90.6 | A | Major actin | P12431 | ACTM_STRPU |
90.1 | A | Major actin | Q9Y701 | ACT1_SUIBO |
92.2 | A | Major actin | P30172 | ACT12_SOLTU |
92.3 | A | Major actin | P81228 | ACT5_SOLTU |
92.0 | A | Major actin | P81229 | ACT8_SOLTU |
92.0 | A | Major actin | P93584 | ACT9_SOLTU |
92.0 | A | Major actin | P93372 | ACT4_TOBAC |
94.5 | A | Major actin | P18602 | ACT3_ARTSX |
91.1 | A | Major actin | Q96482 | ACT1_SOLLC |
91.4 | A | Major actin | Q96484 | ACT3_SOLLC |
91.4 | A | Major actin | P93371 | ACT5_TOBAC |
92.0 | A | Major actin | P93375 | ACT7_TOBAC |
91.4 | A | Major actin | Q96483 | ACT2_SOLLC |
91.1 | A | Major actin | P93587 | ACT1_SOLTU |
91.1 | A | Major actin | P93376 | ACT6_TOBAC |
90.5 | A | Major actin | P93585 | ACT4_SOLTU |
92.6 | A | Major actin | Q03342 | ACT3_ECHGR |
91.7 | A | Major actin | Q92192 | ACT_CALFI |
94.0 | A | Major actin | Q92193 | ACT_CRAVI |
96.9 | A | Major actin | P30170 | ACT10_SOLTU |
97.0 | A | Major actin | Q11212 | ACT_SPOLI |
90.1 | A | Major actin | Q25381 | ACTM_LYTPI |
91.3 | A | Major actin | Q25379 | ACT3_LYTPI |
92.6 | A | Major actin | P10982 | ACT1_ABSGL |
95.5 | A | Major actin | P00544 | FGR_FSVGR |
90.4 | A | Major actin | P10994 | ACTS_PLEWA |
97.6 | A | Major actin | Q39596 | ACT_OXYRB |
97.8 | A | Major actin | P85911 | ACT1_PSEMZ |
96.9 | A | Major actin | Q24733 | ACT_DICVI |
100.0 | G | Major actin | P06396 | GELS_HUMAN |
99.2 | G | Major actin | Q3SX14 | GELS_BOVIN |
99.2 | G | Major actin | P20305 | GELS_PIG |
98.4 | G | Major actin | Q28372 | GELS_HORSE |
97.6 | G | Major actin | P13020 | GELS_MOUSE |
97.6 | G | Major actin | Q68FP1 | GELS_RAT |