scholarly journals The calcium-binding site of human glutamate carboxypeptidase II is critical for dimerization, thermal stability, and enzymatic activity

2018 ◽  
Vol 27 (9) ◽  
pp. 1575-1584 ◽  
Author(s):  
Jakub Ptacek ◽  
Jana Nedvedova ◽  
Michal Navratil ◽  
Barbora Havlinova ◽  
Jan Konvalinka ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Enzymatic Activity ◽  
Binding Site ◽  
Calcium Binding ◽  
Calcium Binding Site ◽  
Glutamate Carboxypeptidase Ii ◽  
Glutamate Carboxypeptidase

scholarly journals A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate

FEBS Open Bio ◽  
2016 ◽  
Vol 6 (5) ◽  
pp. 425-432 ◽  
Author(s):  
Johannes Then ◽  
Ren Wei ◽  
Thorsten Oeser ◽  
André Gerdts ◽  
Juliane Schmidt ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Binding Site ◽  
Polyethylene Terephthalate ◽  
Calcium Binding ◽  
Disulfide Bridge ◽  
Calcium Binding Site

scholarly journals Calcium regulates tertiary structure and enzymatic activity of human endometase/matrilysin-2 and its role in promoting human breast cancer cell invasion

2007 ◽  
Vol 403 (1) ◽  
pp. 31-42 ◽  
Author(s):  
Seakwoo Lee ◽  
Hyun I. Park ◽  
Qing-Xiang Amy Sang
Keyword(s):  
Breast Cancer ◽  
Enzymatic Activity ◽  
Binding Site ◽  
Calcium Binding ◽  
Human Breast Cancer ◽  
Structural Changes ◽  
Human Breast Cancer Cell ◽  
Human Breast ◽  
Calcium Binding Site ◽  
High Affinity

Human MMP-26 (matrix metalloproteinase-26) (also known as endometase or matrilysin-2) is a putative biomarker for human carcinomas of breast, prostate and other cancers of epithelial origin. Calcium modulates protein structure and function and may act as a molecular signal or switch in cells. The relationship between MMPs and calcium has barely been studied and is absent for MMP-26. We have investigated the calcium-binding sites and the role of calcium in MMP-26. MMP-26 has one high-affinity and one low-affinity calcium binding site. High-affinity calcium binding was restored at physiologically low calcium conditions with a calcium-dissociation constant of 63 nM without inducing secondary and tertiary structural changes. High-affinity calcium binding protects MMP-26 against thermal denaturation. Mutants of this site (D165A or E191A) lose enzymatic activity. Low-affinity calcium binding was restored at relatively high calcium concentrations and showed a Kd2 (low-affinity calcium-dissociation constant) value of 120 μM, which was accompanied with the recovery of enzymatic activity reversibly and tertiary structural changes, but without secondary structural rearrangements. Mutations at the low-affinity calcium-binding site (C3 site), K189E or D114A, induced enhanced affinity for the Ca2+ ion or an irreversible loss of enzymatic activity triggered by low-affinity calcium binding respectively. Mutation at non-calcium-binding site (V184D at C2 site) showed that C2 is not a true calcium-binding site. Observations from homology-modelled mutant structures correlated with these experimental results. A human breast cancer cell line, MDA-MB-231, transfected with wild-type MMP-26 cDNA showed a calcium-dependent invasive potential when compared with controls that were transfected with an inactive form of MMP-26 (E209A). Calcium-independent high invasiveness was observed in the K189E mutant MDA-MB-231 cell line.

The structural analysis and the role of calcium binding site for thermal stability in mannanase

Biochimie ◽  
2012 ◽  
Vol 94 (12) ◽  
pp. 2783-2790 ◽  
Author(s):  
Yuya Kumagai ◽  
Kayoko Kawakami ◽  
Takafumi Mukaihara ◽  
Masayo Kimura ◽  
Tadashi Hatanaka
Keyword(s):  
Thermal Stability ◽  
Structural Analysis ◽  
Binding Site ◽  
Calcium Binding ◽  
Calcium Binding Site

A Single Calcium Binding Site Is Crucial for the Calcium-Dependent Thermal Stability of Thermolysin-like Proteases†

Biochemistry ◽  
1998 ◽  
Vol 37 (15) ◽  
pp. 5312-5319 ◽  
Author(s):  
Oene R. Veltman ◽  
Gert Vriend ◽  
Herman J. C. Berendsen ◽  
Bertus Van den Burg ◽  
Gerard Venema ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Binding Site ◽  
Calcium Binding ◽  
Calcium Binding Site ◽  
Calcium Dependent ◽  
Thermal Stability Of
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