Two-dimensional cross correlation analysis of protein unfolding: Portrayal of the thermal denaturation of CMP kinases in the absence and presence of substrates

Schultz, Christian P.; Ba⁁rzu, Octavian; Mantsch, Henry H.
March 2000
AIP Conference Proceedings;2000, Vol. 503 Issue 1, p85
Academic Journal
The functional role of CMP kinases is to regenerate mono-phosphate nucleotides in cells by transferring phosphate residues from tri-phosphorylated nucleotides to monophosphorylated nucleotides. These enzymes possess two binding sites and maintain a highly conserved secondary structure. They are essential for cell survival. Herein we compare the infrared spectra of two similar, but not identical enzymes, the CMP kinases from Escherichia coli and Bacillus subtilis. A two-dimensional cross correlation analysis of the infrared spectra reveals differences in the denaturation behavior of the two proteins. Different secondary structure elements show different time-delayed or advanced unfolding events in the two enzymes. When bound to the active sites, the two nucleotide-substrates CMP and ATP exert a stabilizing effect on the structure of both proteins. The changes observed upon thermal denaturation are different for the two enzymes. Model 2D correlations are used to simulate the different denaturation of the two enzymes. Thermal denaturation and aggregation can be distinguished as two processes separated in time. © 2000 American Institute of Physics.


Related Articles

  • The Stable, Inactive but Reactivable, Unfolding Intermediate of Rat Muscle Sarcoplasmic Reticulum... Bergenhem, Nils C.H.; Lee, Sun-Joo // Journals of Gerontology Series A: Biological Sciences & Medical ;Sep97, Vol. 52A Issue 5, pB240 

    Presents information on a study which investigated guanidinium-chloride-induced inactivation, and reactivation by dilution of the denaturant, of rat muscle sarcoplasmic reticulum CA...+-ATPase enzyme. Methods; Results and discussion; Conclusions.

  • Structural and Functional Characterization of Rabbit and Human l-Gulonate 3-Dehydrogenase. Ishikura, Syuhei; Usami, Noriyuki; Araki, Mayuko; Hara, Akira // Journal of Biochemistry;Mar2005, Vol. 137 Issue 3, p303 

    l-Gulonate 3-dehydrogenase (GDH) catalyzes the NAD+-linked dehydrogenation of l-gulonate into dehydro-l-gulonate in the uronate cycle. In this study, we isolated the enzyme and its cDNA from rabbit liver, and found that the cDNA is identical to that for rabbit lens λ-crystallin except for...

  • Isolation, purification, and characterization of thermophilic laccase from the xerophyte Cereus pterogonus. Gali, Nirmal; Kotteazeth, Srikumar // Chemistry of Natural Compounds;Jul2012, Vol. 48 Issue 3, p451 

    Three laccase temperature isoforms were isolated and purified to homogeneity from the xerophyte plant species Cereus pterogonus. This catalytically active protein exhibited an apparent molecular mass of 137 kDa, 90 kDa, and 43 kDa. Under reducing conditions the enzyme yielded a subunit molecular...

  • Structural Changes in Halophilic and Non-halophilic Proteases in Response to Chaotropic Reagents. Sinha, Rajeshwari; Khare, S. // Protein Journal;Aug2014, Vol. 33 Issue 4, p394 

    Halophilic enzymes have been established for their stability and catalytic abilities under harsh operational conditions. These have been documented to withstand denaturation at high temperature, pH, organic solvents, and chaotropic agents. However, this stability is modulated by salt. The...

  • Using Enzyme Folding to Explore the Mechanism of Therapeutic Touch: A Feasibility Study. Strickland, Mallory L.; Boylan, Helen M. // Journal of Alternative & Complementary Medicine;Jul2010, Vol. 16 Issue 7, p715 

    Objectives: The goal of this research is to design a novel model using protein folding to study Therapeutic Touch, a noncontact form of energy manipulation healing. Presented is a feasibility study suggesting that the denaturation path of ribonuclease A may be a useful model to study the energy...

  • Evidence for proteasome involvement in polyglutamine disease: localization to nuclear inclusions in SCA3/MJD and suppression of polyglutamine aggregation. Chai, Yaohui; Koppenhafer, Stacia L.; Shoesmith, Sarah J.; Perez, Matthew K.; Paulson, Henry L. // Human Molecular Genetics;Apr99, Vol. 8 Issue 4, p673 

    Spinocerebellar ataxia type 3, also known as Machado-Joseph disease (SCA3/MJD), is one of at least eight inherited neurodegenerative diseases caused by expansion of a polyglutamine tract in the disease protein. Here we present two lines of evidence implicating the ubiquitin-proteasome pathway in...

  • Chaperones: A story of thrift unfolds. Baneyx, François; Nannenga, Brent L. // Nature Chemical Biology;Dec2010, Vol. 6 Issue 12, p880 

    The article reports on a new substrate that helps establish DnaK as an unfoldase that needs at least five adenosine triphosphate (ATP) molecules to cause the refolding of one protein. It says that chaperone enzymology has been examined using enzymes denatured chemically or by heat. It adds that...

  • Improved A. faecalis Penicillin Amidase Mutant Retains the Thermodynamic and pH Stability of the Wild Type Enzyme. Yuryev, Ruslan; Kasche, Volker; Ignatova, Zoya; Galunsky, Boris // Protein Journal;Apr2010, Vol. 29 Issue 3, p181 

    Penicillin amidase from Alacaligenes faecalis is an attractive biocatalyst for hydrolysis of penicillin G for production of 6-aminopenicillanic acid, which is used in the synthesis of semi-synthetic β-lactam antibiotics. Recently a mutant of this enzyme with extended C-terminus of the A-chain...

  • A lysozyme unfolding mechanism. Volynskaya, A. V.; Murasheva, S. A.; Skripkin, A. Yu.; Shishkov, A. V. // Doklady Physical Chemistry;Feb2010, Vol. 430 Issue 2, p29 

    The article reports on the study conducted to determine the unfolding mechanism of lysozyme under equilibrium conditions in urea solution using the tritium label method. The refolding kinetics of the protein showed the involvement of two intermediates in the folding pathway. The study revealed...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics