Conformational Dynamics of the GdmHCl-Induced Molten Globule State of Creatine Kinase Monitored by Hydrogen Exchange and Mass Spectrometry

Biochemistry ◽  
2004 ◽  
Vol 43 (17) ◽  
pp. 5045-5054 ◽  
Author(s):  
Hortense Mazon ◽  
Olivier Marcillat ◽  
Eric Forest ◽  
David L. Smith ◽  
Christian Vial
Keyword(s):  
Mass Spectrometry ◽  
Creatine Kinase ◽  
Hydrogen Exchange ◽  
Molten Globule ◽  
Conformational Dynamics ◽  
Molten Globule State

Structural Characterization of the Molten Globule State of Apomyoglobin by Limited Proteolysis and HPLC-Mass Spectrometry†

Biochemistry ◽  
2005 ◽  
Vol 44 (20) ◽  
pp. 7490-7496 ◽  
Author(s):  
Yeoun Jin Kim ◽  
Young A Kim ◽  
Nokyoung Park ◽  
Hyeon S. Son ◽  
Kwang S. Kim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Structural Characterization ◽  
Molten Globule ◽  
Limited Proteolysis ◽  
Molten Globule State

The Conformational Dynamics of a Metastable Serpin Studied by Hydrogen Exchange and Mass Spectrometry

Biochemistry ◽  
2006 ◽  
Vol 45 (21) ◽  
pp. 6561-6569 ◽  
Author(s):  
Yuko Tsutsui ◽  
Lu Liu ◽  
Anne Gershenson ◽  
Patrick L. Wintrode
Keyword(s):  
Mass Spectrometry ◽  
Hydrogen Exchange ◽  
Conformational Dynamics

Probing subtle differences in the hydrogen exchange behavior of variants of the human α-lactalbumin molten globule using mass spectrometry

2001 ◽  
Vol 311 (4) ◽  
pp. 909-919 ◽  
Author(s):  
Alexander M Last ◽  
Brenda A Schulman ◽  
Carol V Robinson ◽  
Christina Redfield
Keyword(s):  
Mass Spectrometry ◽  
Hydrogen Exchange ◽  
Molten Globule

Effects of lactic acid and NaCl on creatine kinase from rabbit muscle

2003 ◽  
Vol 81 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Hong-Min Tang ◽  
Wen-Bin Ou ◽  
Hai-Meng Zhou
Keyword(s):  
Creatine Kinase ◽  
Lactic Acid ◽  
Gel Electrophoresis ◽  
Molten Globule ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Rabbit Muscle ◽  
Molten Globule State ◽  
Induced Folding ◽  
Native Polyacrylamide Gel Electrophoresis

The lactic acid induced unfolding and the salt-induced folding of creatine kinase (CK) were studied by enzyme activity, fluorescence emission spectra, circular dichroism spectra, and native polyacrylamide gel electrophoresis. The results showed that the kinetics of CK inactivation was a monophase process. Lactic acid caused inactivation and unfolding of CK with no aggregation during CK denaturation. The unfolding of the whole molecule and the inactivation of CK in solutions of different concentration of lactic acid were compared. Much lower lactic acid concentration values were required to bring about inactivation than were required to produce significant conformational changes of the enzyme molecule. At higher concentrations of lactic acid (more than 0.2 mM) the CK dimers were partially dissociated, as proved by native polyacrylamide gel electrophoresis. NaCl induced the molten globule state with a compact structure after CK was denatured with 0.8 mM lactic acid, and the increasing of anions led to a tight side-chain. The above results suggest that the effect of lactic acid differed from that of other denaturants such as guanidine hydrochloride, HCl, or urea during CK folding, and the molten globule state indicates that intermediates exist during CK folding.Key words: lactic acid, creatine kinase, salt-induced, unfolding, molten globule state.

Structure and stability of the molten globule state of guinea pig .alpha.-lactalbumin: A hydrogen exchange study

Biochemistry ◽  
1993 ◽  
Vol 32 (21) ◽  
pp. 5681-5691 ◽  
Author(s):  
Chia Lin Chyan ◽  
Claire Wormald ◽  
Christopher M. Dobson ◽  
Philip A. Evans ◽  
Jean Baum
Keyword(s):  
Guinea Pig ◽  
Hydrogen Exchange ◽  
Molten Globule ◽  
Molten Globule State ◽  
Structure And Stability ◽  
Exchange Study ◽  
Alpha Lactalbumin

Noncovalent binding of a cyclic peptide inhibitor to the peptidyl-prolyl isomerase Pin1, explored by hydrogen exchange mass spectrometry

2015 ◽  
Vol 93 (1) ◽  
pp. 44-50
Author(s):  
Modupeola A. Sowole ◽  
Brendan T. Innes ◽  
Mahasilu Amunugama ◽  
David W. Litchfield ◽  
Christopher J. Brandl ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Hydrogen Exchange ◽  
Cyclic Peptide ◽  
Conformational Dynamics ◽  
High Specificity ◽  
Prolyl Isomerase ◽  
Peptidyl Prolyl Isomerase ◽  
X Ray ◽  
Exchange Mass Spectrometry ◽  
Ppiase Domain

Pin1 is a peptidyl-prolyl isomerase (PPIase) that plays a central role in eukaryotic cell cycle regulation, making this protein an interesting target for cancer therapy. Pin1 exhibits high specificity for substrates where proline is preceded by phosphoserine or phosphothreonine. The protein comprises an N-terminal WW (tryptophan–tryptophan) domain and a C-terminal PPIase domain. The cyclic peptide [CRYPEVEIC] (square brackets are used to denote the cyclic structure) represents a lead compound for a new class of nonphosphorylated Pin1 inhibitors. Unfortunately, it has not been possible thus far to characterize the Pin1–[CRYPEVEIC] complex by X-ray crystallography. Thus, the exact binding mode remains unknown. The current work employs hydrogen/deuterium exchange mass spectrometry for gaining insights into the Pin1–[CRYPEVEIC] interactions. The WW domain shows extensive conformational dynamics, both in the presence and in the absence of ligand. In contrast, profound changes in deuteration kinetics are observed in the PPIase domain after the addition of [CRYPEVEIC]. The secondary structure elements β2, α3, and α4 exhibit markedly reduced deuteration, consistent with their postulated involvement in ligand binding. Unexpectedly, [CRYPEVEIC] destabilizes the range of residues 61–86, a segment that comprises basic side chains that normally interact with the substrate phosphate. This destabilization is likely caused by steric clashes with Y3 or E5 of the inhibitor. Ligand-induced destabilization has previously been reported for a few other proteins, but effects of this type are not very common. Our findings suggest that future crystallization trials on Pin1 variants deleted for residues in the 61–86 range might provide a path towards high-resolution X-ray structures of Pin1 bound to cyclic peptide inhibitors.

scholarly journals Hydrogen exchange study of folding intermediate and molten globule state of canine milk lysozyme

2000 ◽  
Vol 40 (supplement) ◽  
pp. S160
Author(s):  
Y. Kobashigawa ◽  
M Mizuguchi ◽  
M. Demura ◽  
T. Koshiba ◽  
S. Tsuda ◽  
...  
Keyword(s):  
Hydrogen Exchange ◽  
Molten Globule ◽  
Folding Intermediate ◽  
Molten Globule State ◽  
Exchange Study
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