Effect of antisense peptide binding on the dimerization of human cystatin C--gel electrophoresis and molecular modeling studies.

K, Stachowiak; Rodziewicz‐Motowidło, Sylwia; Sosnowska, Renata; Kasprzykowski, Franciszek; Łankiewicz, Leszek; Grubb, Anders; Grzonka, Zbigniew

doi:10.18388/abp.2004_3607

Cited by 2 publications

(2 citation statements)

References 33 publications

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“…The electrophoretic mobility of all the studied proteins in an agarose gel was also checked. This technique had earlier been shown to allow simultaneous observation of different oligomeric forms of cystatin C based on differences in the mobility of protein dimers and monomers (Ekiel & Abrahamson, 1996;Stachowiak et al, 2004). As expected, substitution of the neutral valine residue by a polar but not charged asparagine did not change the position of the protein band comparing to wild type cystatin C (Fig.…”

Section: Physicochemical Characterization Of Hcc Variantssupporting

confidence: 63%

Governing the monomer-dimer ratio of human cystatin c by single amino acid substitution in the hinge region.

Szymańska¹,

Radulska²,

Czaplewska³

et al. 2009

Acta Biochim Pol

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Three dimensional domain swapping is one of the mechanisms involved in formation of insoluble aggregates of some amyloidogenic proteins. It has been proposed that proteins able to swap domains may share some common structural elements like conformationally constrained flexible turns/loops. We studied the role of loop L1 in the dimerization of human cystatin C using mutational analysis. Introduction of turn-favoring residues such as Asp or Asn into the loop sequence (in position 57) leads to a significant reduction of the dimer fraction in comparison with the wild type protein. On the other hand, introduction of a proline residue in position 57 leads to efficient dimer formation. Our results confirm the important role of the loop L1 in the dimerization process of human cystatin C and show that this process can be to some extent governed by single amino acid substitution.

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Section: Physicochemical Characterization Of Hcc Variantssupporting

confidence: 63%

Governing the monomer-dimer ratio of human cystatin c by single amino acid substitution in the hinge region.

Szymańska¹,

Radulska²,

Czaplewska³

et al. 2009

Acta Biochim Pol

Self Cite

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show abstract

“…5); the amounts of cystatin C protein were equivalent in cells grown on plastic compared to those on Matrigel. An additional 25 kDA band was detected in the media of both cell lines; this band may represent cystatin C dimerization (38,39) or different glycoforms of the inhibitor. Various glycoforms of cystatin C similar in size to our 25 kDa band were also detected in media conditioned by hippocampal stem cells (40).…”

Section: Immunolocalization Of Cathepsins B and L Co-immunolocalizatmentioning

confidence: 96%

Matrigel influences morphology and cathepsin B distribution of prostate cancer PC3 cells

Gopal

Rehman²,

Chadha³

et al. 2006

Oncol Rep

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Increases in expression and activity of matrixdegrading enzymes such as the cysteine proteinases cathepsins B and L, and abnormal levels of their inhibitors, the cystatins, are associated with tumor cell invasion and metastasis. Environmental conditions have been shown to be causative factors in the development of a metastatic/invasive phenotype. We hypothesized that cell-matrix interactions affect the expression and activity of cathepsins B and L and their inhibitors in the prostate cancer cell lines, PC3 and DU145. To test this possibility, PC3 and DU145 were plated on uncoated surfaces or on surfaces coated with the reconstituted basement membrane, Matrigel. The cells were analyzed for cathepsins B and L immunolocalization, protein expression and activity 48 h after plating. Our data demonstrated that cathepsins B and L displayed a distinct punctate distribution with little colocalization; individual cells displayed a predominant staining for one or the other enzyme. Cathepsin B had a perinuclear distribution in PC3 grown on uncoated surfaces but a more peripheral staining in PC3 plated on Matrigel. Localization of cathepsin L remained predominantly perinuclear regardless of the plating surface. In addition to the translocation of cathepsin B from a perinuclear distribution to the cell periphery, growth of PC3 on Matrigel shifted cathepsin B activity from the cell extract to the media. There were no significant changes in cathepsins B and L immunolocalization or activity in DU145 with regard to plating surfaces. Likewise, the activity of endogenous cysteine proteinase inhibitors (CPIs) and protein expression of cystatin C remained unchanged in both cell lines. In conclusion, the interaction of PC3 prostate cancer cells with extracellular matrix components affects the distribution of cathepsin B protein and activity.

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Effect of antisense peptide binding on the dimerization of human cystatin C--gel electrophoresis and molecular modeling studies.

Cited by 2 publications

References 33 publications

Governing the monomer-dimer ratio of human cystatin c by single amino acid substitution in the hinge region.

Governing the monomer-dimer ratio of human cystatin c by single amino acid substitution in the hinge region.

Matrigel influences morphology and cathepsin B distribution of prostate cancer PC3 cells

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