A new folding intermediate of apomyoglobin from Aplysia limacina: stepwise formation of a molten globule.


Abstract

Apomyoglobin from Aplysia limacina (al-apoMb), despite having only 20 % sequence identity with the more commonly studied mammalian globins (m-apoMbs), properties which result in an increased number of hydrophobic contacts and a loss of most internal salt bridges, shares a number of features of their folding profiles. We show here that it contains an unusually stable core which resists unfolding even at 70 degrees C. The equilibrium intermediate (I(T)) at this high temperature is distinct from the acid unfolded state I(A) which has many properties in common with the acid intermediate observed for the mammalian apoproteins (I(AGH)). It contains a smaller amount of secondary structure (27 % alpha-helical instead of 35 %) and is more highly solvated as evidenced from its fluorescence spectrum (lambda(max)=344 nm instead of 338 nm). Its stability is greatly increased (DeltaDeltaG(w)=-6.75 kcal mol(-1)) in the presence of high salt (2 M KCl), lending support to the view that hydrophobic interactions are responsible for its stability. Kinetic data show classical two-state kinetics between I(A) and the folded state both in the presence and absence of salt. Both I(A) and I(T) can be populated within the dead time of the stopped-flow apparatus, since initiating the refolding reaction from I(T) or I(A) rather than the completely unfolded state does not affect the observed refolding time-course. Our conclusion is that al-apoMb, as other "apo" proteins (including for example alpha-lactalbumin in the absence of Ca(2+)), may be described as "uncoupled" with an unusually high and exploitable tendency to populate partially folded states. Study holds ProTherm entries: 8601, 8602 Extra Details: apomyoglobin; folding intermediate; cooperativity of folding

Submission Details

ID: da4yUUdm3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:36 p.m.

Version: 1

Publication Details
Staniforth RA;Giannini S;Bigotti MG;CutruzzolĂ  F;Travaglini-Allocatelli C;Brunori M,J. Mol. Biol. (2000) A new folding intermediate of apomyoglobin from Aplysia limacina: stepwise formation of a molten globule. PMID:10764586
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
1MBA 1990-01-15 1.6 APLYSIA LIMACINA MYOGLOBIN. CRYSTALLOGRAPHIC ANALYSIS AT 1.6 ANGSTROMS RESOLUTION
2FAL 1993-10-31 1.8 X-RAY CRYSTAL STRUCTURE OF FERRIC APLYSIA LIMACINA MYOGLOBIN IN DIFFERENT LIGANDED STATES
5MBA 1992-07-15 1.9 BINDING MODE OF AZIDE TO FERRIC APLYSIA LIMACINA MYOGLOBIN. CRYSTALLOGRAPHIC ANALYSIS AT 1.9 ANGSTROMS RESOLUTION
1DM1 2000-06-21 1.99 2.0 A CRYSTAL STRUCTURE OF THE DOUBLE MUTANT H(E7)V, T(E10)R OF MYOGLOBIN FROM APLYSIA LIMACINA
2FAM 1993-10-31 2.0 X-RAY CRYSTAL STRUCTURE OF FERRIC APLYSIA LIMACINA MYOGLOBIN IN DIFFERENT LIGANDED STATES
4MBA 1990-01-15 2.0 APLYSIA LIMACINA MYOGLOBIN. CRYSTALLOGRAPHIC ANALYSIS AT 1.6 ANGSTROMS RESOLUTION
3MBA 1990-01-15 2.0 APLYSIA LIMACINA MYOGLOBIN. CRYSTALLOGRAPHIC ANALYSIS AT 1.6 ANGSTROMS RESOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Globin P02210 GLB_APLLI