Unfolding and refolding of a type kappa immunoglobulin light chain and its variable and constant fragments.


Abstract

By limited proteolysis of a type kappa immunoglobulin light chain (Oku) with clostripain, both the VL and CL fragments were obtained with a high yield. The unfolding and refolding by guanidine hydrochloride of light chain Oku and its VL and CL fragments were studied at pH 7.5 and 25 degrees C with circular dichroism and tryptophyl fluorescence. The concentration of guanidine hydrochloride at the midpoint of the unfolding curve was 1.2 M for the VL fragment and 0.9 M for the CL fragment. As in the case of the CL fragment of light chain Nag (type lambda) [Goto, Y., & Hamaguchi, K. (1982) J. Mol. Biol. 156, 891-910], the unfolding and refolding kinetics of the CL fragment could be explained in principle on the basis of the three-species mechanism U1 in equilibrium U2 in equilibrium N, where N is native protein and U1 and U2 are the slow-folding and fast-folding species, respectively, of unfolded protein. The unfolding and refolding kinetics of the VL(Oku) fragment were described by a single exponential term. Double-jump experiments, however, showed that two forms of the unfolding molecule exist. The equilibrium and kinetics of unfolding of light chain Oku were explained by the sum of those of the VL and CL fragments. On the other hand, the refolding kinetics of light chain Oku were greatly different from the sum of those of the VL and CL fragments.(ABSTRACT TRUNCATED AT 250 WORDS) Study holds ProTherm entries: 3907, 3908 Extra Details: unfolding and refolding kinetics; three-species mechanism;,double-jump experiment; isolated domains; slow phase

Submission Details

ID: r7gKpvyR

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:23 p.m.

Version: 1

Publication Details
Tsunenaga M;Goto Y;Kawata Y;Hamaguchi K,Biochemistry (1987) Unfolding and refolding of a type kappa immunoglobulin light chain and its variable and constant fragments. PMID:3120770
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
2Q20 2008-04-08 1.3 Structure of the germline Vk1 O18/O8 light chain variable domain homodimer
1EEQ 2001-02-01 1.5 M4L/Y(27D)D/T94H Mutant of LEN
3CDF 2008-09-02 1.53 kI O18/O8 Y87H immunoglobulin light chain variable domain
3CDC 2008-09-02 1.53 kI O18/O8 N34I/Y87H immunoglobulin light chain variable domain
1EEU 2001-02-03 1.6 M4L/Y(27D)D/Q89D/T94H mutant of LEN
1EFQ 2001-02-09 1.6 Q38D mutant of LEN
4L1H 2014-06-04 1.68 Bence-Jones immunoglobulin REI variable portion with seven point mutations
1BWW 1998-10-07 1.7 BENCE-JONES IMMUNOGLOBULIN REI VARIABLE PORTION, T39K MUTANT
1QAC 2000-02-23 1.8 CHANGE IN DIMERIZATION MODE BY REMOVAL OF A SINGLE UNSATISFIED POLAR RESIDUE
1B0W 1998-11-16 1.8 Structural comparison of amyloidogenic light chain dimer in two crystal forms with nonamyloidogenic counterparts
1EK3 2001-03-06 1.9 KAPPA-4 IMMUNOGLOBULIN VL, REC
1WTL 1994-11-01 1.9 COMPARISON OF CRYSTAL STRUCTURES OF TWO HOMOLOGOUS PROTEINS: STRUCTURAL ORIGIN OF ALTERED DOMAIN INTERACTIONS IN IMMUNOGLOBULIN LIGHT CHAIN DIMERS
1LVE 1998-01-21 1.95 STRUCTURE OF THE VARIABLE DOMAIN OF HUMAN IMMUNOGLOBULIN K-4 LIGHT CHAIN LEN
1REI 1976-05-19 2.0 THE MOLECULAR STRUCTURE OF A DIMER COMPOSED OF THE VARIABLE PORTIONS OF THE BENCE-JONES PROTEIN REI REFINED AT 2.0 ANGSTROMS RESOLUTION
3LVE 1999-05-18 2.0 LEN Q38E MUTANT: A DOMAIN FLIP FROM A SINGLE AMINO ACID SUBSTITUTION
5LVE 2000-02-18 2.0 STRUCTURE OF THE VARIABLE DOMAIN OF HUMAN IMMUNOGLOBULIN K-4 LIGHT CHAIN LEN
1QP1 1999-06-17 2.06 KAPPA VARIABLE LIGHT CHAIN
1JV5 2002-01-09 2.2 Anti-blood group A Fv
4LVE 1999-05-18 2.3 LEN K30T MUTANT: A DOMAIN FLIP AS A RESULT OF A SINGLE AMINO ACID SUBSTITUTION
3CDY 2008-09-02 2.43 AL-09 H87Y, immunoglobulin light chain variable domain
2LVE 1999-05-18 2.7 RECOMBINANT LEN
1AR2 1997-11-12 2.8 DISULFIDE-FREE IMMUNOGLOBULIN FRAGMENT
4K07 2013-10-30 2.83 Crystal structure of the amyloid-forming immunoglobulin AL-103 cis-proline 95 mutant
5XP1 2017-08-02 2.88 Structure of monomeric mutant of REI immunoglobulin light chain variable domain crystallized at pH 6

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Immunoglobulin kappa variable 1D-33 P01593 KVD33_HUMAN
100.0 Immunoglobulin kappa variable 1D-33 P01594 KV133_HUMAN
99.0 Immunoglobulin kappa variable 4-1 P06312 KV401_HUMAN