Hierarchical protein folding: asymmetric unfolding of an insulin analogue lacking the A7-B7 interchain disulfide bridge.


Abstract

The landscape paradigm of protein folding can enable preferred pathways on a funnel-like energy surface. Hierarchical preferences may be manifest as a nonrandom pathway of disulfide pairing. Stepwise stabilization of structural subdomains among on-pathway intermediates is proposed to underlie the disulfide pathway of proinsulin and related molecules. Here, effects of pairwise serine substitution of insulin's exposed interchain disulfide bridge (Cys(A7)-Cys(B7)) are characterized as a model of a late intermediate. Untethering cystine A7-B7 in an engineered monomer causes significantly more marked decreases in the thermodynamic stability and extent of folding than occur on pairwise substitution of internal cystine A6-A11 [Weiss, M. A., Hua, Q. X., Jia, W., Chu, Y. C., Wang, R. Y., and Katsoyannis, P. G. (2000) Biochemistry 39, 15429-15440]. Although substantially disordered and without significant biological activity, the untethered analogue contains a molten subdomain comprising cystine A20-B19 and a native-like cluster of hydrophobic side chains. Remarkably, A and B chains make unequal contributions to this folded moiety; the B chain retains native-like supersecondary structure, whereas the A chain is largely disordered. These observations suggest that the B subdomain provides a template to guide folding of the A chain. Stepwise organization of insulin-like molecules supports a hierarchic view of protein folding. Study holds ProTherm entries: 12271 Extra Details: funnel-like energy surface; disulfide pathway; supersecondary structure; hierarchic

Submission Details

ID: ApDMtf6N3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:43 p.m.

Version: 1

Publication Details
Hua QX;Nakagawa SH;Jia W;Hu SQ;Chu YC;Katsoyannis PG;Weiss MA,Biochemistry (2001) Hierarchical protein folding: asymmetric unfolding of an insulin analogue lacking the A7-B7 interchain disulfide bridge. PMID:11591149
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
1BZV 1998-11-04T00:00:00+0000 0 [D-ALAB26]-DES(B27-B30)-INSULIN-B26-AMIDE A SUPERPOTENT SINGLE-REPLACEMENT INSULIN ANALOGUE, NMR, MINIMIZED AVERAGE STRUCTURE
1BZV 1998-11-04T00:00:00+0000 0 [D-ALAB26]-DES(B27-B30)-INSULIN-B26-AMIDE A SUPERPOTENT SINGLE-REPLACEMENT INSULIN ANALOGUE, NMR, MINIMIZED AVERAGE STRUCTURE
3WS3 2014-02-28T00:00:00+0000 2.33 Crystal Structure of H-2D in complex with an insulin derived peptide
6CE7 2018-02-11T00:00:00+0000 7.4 Insulin Receptor ectodomain in complex with one insulin molecule
6CE9 2018-02-11T00:00:00+0000 4.3 Insulin Receptor ectodomain in complex with two insulin molecules
6CEB 2018-02-11T00:00:00+0000 4.7 Insulin Receptor ectodomain in complex with two insulin molecules - C1 symmetry
1APH 1992-10-30T00:00:00+0000 2.0 CONFORMATIONAL CHANGES IN CUBIC INSULIN CRYSTALS IN THE PH RANGE 7-11
1APH 1992-10-30T00:00:00+0000 2.0 CONFORMATIONAL CHANGES IN CUBIC INSULIN CRYSTALS IN THE PH RANGE 7-11
1BPH 1992-10-30T00:00:00+0000 2.0 CONFORMATIONAL CHANGES IN CUBIC INSULIN CRYSTALS IN THE PH RANGE 7-11
1BPH 1992-10-30T00:00:00+0000 2.0 CONFORMATIONAL CHANGES IN CUBIC INSULIN CRYSTALS IN THE PH RANGE 7-11

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
547.8 B,D,F,H,J,L Insulin P01338 INS2_BATSP
550.2 B,D,F,H,J,L Insulin P68987 INS_PETMA
550.2 B,D,F,H,J,L Insulin P68988 INS_LAMFL
555.6 B,D,F,H,J,L Insulin C0HJI7 INS1_HUSDA
553.8 B,D,F,H,J,L Insulin C0HJI3 INS2_KATPE
553.8 B,D,F,H,J,L Insulin P09476 INS_ATRSP
553.8 B,D,F,H,J,L Insulin Q9W7R2 INS_VERMO
553.8 B,D,F,H,J,L Insulin P09477 INS_PLAFE
553.8 B,D,F,H,J,L Insulin P04667 INS_ONCKE
555.6 B,D,F,H,J,L Insulin P01340 INS_KATPE
558.6 B,D,F,H,J,L Insulin P83770 INSL_VIGUN
555.6 B,D,F,H,J,L Insulin P12707 INS2_XENLA
577.2 B,D,F,H,J,L Insulin C0HJI8 INS2_HUSDA
579.6 B,D,F,H,J,L Insulin P21563 INS_RODSP
600.0 B,D,F,H,J,L Insulin P81423 INS_ACIGU
557.4 B,D,F,H,J,L Insulin P12706 INS1_XENLA
577.8 B,D,F,H,J,L Insulin P69048 INS_TRASC
577.8 B,D,F,H,J,L Insulin P69047 INS_TRADO
577.8 B,D,F,H,J,L Insulin P67969 INS_STRCA
577.8 B,D,F,H,J,L Insulin P67968 INS_MELGA
577.8 B,D,F,H,J,L Insulin P67970 INS_CHICK
1149.6000000000001 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01323 INS2_RAT
1149.6000000000001 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01326 INS2_MOUSE
578.4 B,D,F,H,J,L Insulin P51463 INS_SELRF
578.4 B,D,F,H,J,L Insulin P68243 INS_CAIMO
578.4 B,D,F,H,J,L Insulin P68245 INS_ANSAN
578.4 B,D,F,H,J,L Insulin P01333 INS_ANAPL
1143.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P18109 INS_DIDVI
559.8 B,D,F,H,J,L Insulin P67971 INS_SAISC
559.8 B,D,F,H,J,L Insulin P67972 INS_AOTTR
1122.6 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01320 INS_CAMDR
1131.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01324 INS_ACOCA
600.0 B,D,F,H,J,L Insulin P01318 INS_SHEEP
1171.2 A,B,C,D,E,F,G,H,I,J,K,L Insulin Q62587 INS_PSAOB
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01315 INS_PIG
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P67974 INS_PHYMC
1171.2 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01310 INS_HORSE
600.0 B,D,F,H,J,L Insulin P06306 INS_FELCA
600.0 B,D,F,H,J,L Insulin P01319 INS_CAPHI
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01321 INS_CANLF
1143.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01317 INS_BOVIN
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P67973 INS_BALPH
1143.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01314 INS_BALBO
1180.2 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01311 INS_RABIT
1180.2 A,B,C,D,E,F,G,H,I,J,K,L Insulin Q91XI3 INS_ICTTR
1151.4000000000003 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01313 INS_CRILO
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin Q8HXV2 INS_PONPY
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P30410 INS_PANTR
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P30406 INS_MACFA
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01308 INS_HUMAN
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin Q6YK33 INS_GORGO
1143.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P01316 INS_ELEMA
1200.0 A,B,C,D,E,F,G,H,I,J,K,L Insulin P30407 INS_CHLAE
600.0 B,D,F,H,J,L Insulin F8WCM5 INSR2_HUMAN
543.0 A,C,E,G,I,K Insulin P01327 INS_CHICH
571.2 A,C,E,G,I,K Insulin P01322 INS1_RAT
571.2 A,C,E,G,I,K Insulin P01325 INS1_MOUSE