Structure and properties of K141E mutant of small heat shock protein HSP22 (HspB8, H11) that is expressed in human neuromuscular disorders

Kim, Maria V.; Kasakov, Alexei S.; Seit-Nebi, Alim S.; Marston, Steven B.; Gusev, Nikolai B.

doi:10.1016/j.abb.2006.07.014

Cited by 39 publications

(63 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Titration of the wild-type HspB6 and HspB8 and of their Cys-mutants with bis-ANS in a wide concentration range was accompanied by a linear increase of fluorescence (data not shown). This correlates with our earlier published data and indicates the presence of a large number of low-affinity binding sites on the surface of these proteins (Kasakov et al 2007;Kim et al 2006). In all cases, the difference of the titration curves of the wild-type proteins and their Cysmutants was minimal thus indicating that mutations introduced do not dramatically affect the hydrophobic properties of sHsp.…”

Section: Resultssupporting

confidence: 78%

“…For instance, mutation R120G results in the change of oligomeric structure, chaperone-like activity, and intersubunit interaction of αB-crystallin (Kumar et al 1999;Perng et al 1999;Simon et al 2007;Michiel et al 2009;Bova et al 1999). Similar effects were observed in the case of HspB8 where mutations K137E and/or K141E were also accompanied by significant changes in the structure and properties (Kasakov et al 2007;Kim et al 2006). This difference can be due to the fact that positions occupied by conservative Arg or Lys residues (Arg120 of αB-crystallin and Lys141 of HspB8) are located inside of the sHsp monomer and are involved in many intra-and intermonomer contacts (Michiel et al 2009;Berengian et al 1997Berengian et al , 1999Simon et al 2007).…”

Section: Discussionsupporting

confidence: 62%

“…In the course of the size-exclusion chromatography, HspB8 seems to be present in the form of an equilibrium mixture of monomers and dimers (Kasakov et al 2007;Kim et al 2006). Therefore, elution volume depends on the quantity of protein loaded on the column (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…4c; Kasakov et al 2007;Kim et al 2006). This might indicate that under conditions of chromatography, HspB8 is presented in the form of an equilibrium mixture of monomer and dimer and this equilibrium depends on the protein concentration.…”

Section: Spectroscopic Investigation Of Oxidized Cys-mutants Of Humanmentioning

confidence: 99%

See 3 more Smart Citations

The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins

Mymrikov

Bukach

Seit-Nebi

et al. 2010

Cell Stress and Chaperones

Self Cite

View full text Add to dashboard Cite

Human αB-crystallin and small heat shock proteins HspB6 and HspB8 were mutated so that all endogenous Cys residues were replaced by Ser and the single Cys residue was inserted in a position homologous to that of Cys137 of human HspB1, i.e. in a position presumably located in the central part of β7 strand of the α-crystallin domain. The secondary, tertiary, and quaternary structures of thus obtained Cys-mutants as well as their chaperone-like activity were similar to those of their wildtype counterparts. Mild oxidation of Cys-mutants leads to formation of disulfide bond crosslinking neighboring monomers thus indicating participation of the β7 strand in intersubunit interaction. Oxidation weakly affects the secondary and tertiary structure, does not affect the quaternary structure of αB-crystallin and HspB6, and shifts equilibrium between monomer and dimer of HspB8 towards dimer formation. It is concluded that the β7 strand participates in the intersubunit interaction of four human small heat shock proteins (αB-crystallin, HspB1, HspB6, HspB8) having different structure of β2 strand of α-crystallin domain and different length and composition of variable N-and C-terminal tails.

show abstract

Section: Resultssupporting

confidence: 78%

Section: Discussionsupporting

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Section: Spectroscopic Investigation Of Oxidized Cys-mutants Of Humanmentioning

confidence: 99%

See 2 more Smart Citations

The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins

Mymrikov

Bukach

Seit-Nebi

et al. 2010

Cell Stress and Chaperones

Self Cite

View full text Add to dashboard Cite

show abstract

“…This region was shown to be critical for the homo-dimer formation of several sHSPs, including HspB8 and HspB1, and possibly also for the hetero-dimer formation (Mymrikov et al 2010). The mutation K141E in HspB8 resulted in significant changes in the protein structure and properties that affected also the intersubunit contacts (Kim et al 2006;Kasakov et al 2007). A similar situation was found for the R120G mutation in HspB5 (references in Mymrikov et al 2010).…”

Section: Discussionsupporting

confidence: 48%

Abnormal interaction of motor neuropathy-associated mutant HspB8 (Hsp22) forms with the RNA helicase Ddx20 (gemin3)

Sun

Fontaine

Hoppe

et al. 2010

Cell Stress and Chaperones

View full text Add to dashboard Cite

A number of missense mutations in the two related small heat shock proteins HspB8 (Hsp22) and HspB1 (Hsp27) have been associated with the inherited motor neuron diseases (MND) distal hereditary motor neuropathy and Charcot-Marie-Tooth disease. HspB8 and HspB1 interact with each other, suggesting that these two etiologic factors may act through a common biochemical mechanism. However, their role in neuron biology and in MND is not understood. In a yeast two-hybrid screen, we identified the DEAD box protein Ddx20 (gemin3, DP103) as interacting partner of HspB8. Using co-immunoprecipitation, chemical cross-linking, and in vivo quantitative fluorescence resonance energy transfer, we confirmed this interaction. We also show that the two disease-associated mutant HspB8 forms have abnormally increased binding to Ddx20. Ddx20 itself binds to the survival-of-motor-neurons protein (SMN protein), and mutations in the SMN1 gene cause spinal muscular atrophy, another MND and one of the most prevalent genetic causes of infant mortality. Thus, these protein interaction data have linked the three etiologic factors HspB8, HspB1, and SMN protein, and mutations in any of their genes cause the various forms of MND. Ddx20 and SMN protein are involved in spliceosome assembly and pre-mRNA processing. RNase treatment affected the interaction of the mutant HspB8 with Ddx20 suggesting RNA involvement in this interaction and a potential role of HspB8 in ribonucleoprotein processing.

show abstract

Molecular Biology of Small Heat Shock Proteins associated with Peripheral Neuropathies

Holmgren

Bouhy

Timmerman

2012

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Heat‐shock proteins (HSPs) are molecular chaperones that protect the cell from various types of stress. Although regulated by stress, some of these proteins are constitutively expressed and responsible for quality control and protein folding. Based on their molecular weight, the HSP family mainly consists of two groups: large and small HSPs. The large HSPs require adenosine triphosphate (ATP) for their functioning, whereas the small HSPs (HSPBs) are ATP independent. The latter bind improperly folded proteins and assist in the targeting process for refolding or degradation. These HSPBs are not only molecular chaperones but they are also involved in many essential cellular processes such as apoptosis, autophagy, splicing, cytoskeleton dynamics and neuronal survival. This review focusses on the small HSPs HSPB1, HSPB3 and HSPB8, as mutations in these proteins are causative for Charcot–Marie–Tooth (CMT) disease and distal hereditary motor neuropathies (dHMN). Moreover, this review discusses the functional consequences of these mutations and their role in the length‐dependent neurodegeneration typical of CMT diseases. Key Concepts: The molecular signature of the small HSPs is the conserved α‐crystallin domain. Mutations in HSPB1, HSPB3 and HSPB8 are associated with dHMN and CMT subtypes. Mutant HSPB1 causes instability of the cytoskeletal elements and therefore disorganisation of the neuronal cytoskeleton could underlie the peripheral neuropathies. Mutant HSPB8 may induce pathomechanisms leading to CMT diseases through a loss of autophagy function, rather than a toxic gain of function. The HSPB3 protein differs considerably from the other members regarding sequence similarity such as the poor conservation in the N ‐terminal region and the very short C ‐terminal domain. It remains fragmentary how mutations in these small HSPs initiate a molecular cascade leading to progressive axonal degeneration.

show abstract

Structure and properties of K141E mutant of small heat shock protein HSP22 (HspB8, H11) that is expressed in human neuromuscular disorders

Cited by 39 publications

References 34 publications

The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins

The pivotal role of the β7 strand in the intersubunit contacts of different human small heat shock proteins

Abnormal interaction of motor neuropathy-associated mutant HspB8 (Hsp22) forms with the RNA helicase Ddx20 (gemin3)

Molecular Biology of Small Heat Shock Proteins associated with Peripheral Neuropathies

Contact Info

Product

Resources

About