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p30, a novel protein target of mouse calcyclin (S100A6)
1Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
Abstract:
A novel protein target of mouse calcyclin (S100A6) was detected by a gel overlay method with 125I-labelled calcyclin. Interaction of calcyclin with its 30 kDa target protein (p30) present in Ehrlich ascites tumour (EAT) cells depended on the presence of Ca2+ ions. The binding of p30, evidenced by the reaction with 125I-labelled calcyclin, was found to be of higher affinity than the binding between mouse calcyclin and annexin II or glyceraldehyde-3-phosphate dehydrogenase. Examination of tissue extracts by the gel overlay method has shown that p30 is present not only in the EAT cells but also in mouse brain and spleen. This novel target protein of mouse calcyclin was purified to homogeneity from EAT cells by means of Phenyl-Sepharose chromatography, affinity chromatography and CM-cellulose chromatography. Purified p30 was digested with alpha-chymotrypsin and a partial amino acid sequence of one of the resulting peptides was established. A database search analysis revealed that the sequence is unique, with a similarity of less than 55% to any other known protein sequence.
Insights
Researchers identified a new mouse calcyclin (S100A6) binding protein (p30) in tumor cells. This calcium-dependent interaction shows high affinity, and the novel protein is found in various mouse tissues.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Calcyclin (S100A6) is a calcium-binding protein involved in various cellular processes.
- Identifying novel protein interactions is crucial for understanding protein function and cellular mechanisms.
Purpose of the Study:
- To identify and characterize novel protein targets of mouse calcyclin.
- To investigate the properties and distribution of a newly discovered calcyclin-binding protein.
Main Methods:
- Gel overlay assay using 125I-labelled calcyclin to detect protein interactions.
- Protein purification using Phenyl-Sepharose, affinity, and CM-cellulose chromatography.
- Peptide generation via alpha-chymotrypsin digestion and partial amino acid sequencing.
Main Results:
- A novel 30 kDa protein (p30) interacting with mouse calcyclin was identified in Ehrlich ascites tumour (EAT) cells.
- The interaction between calcyclin and p30 is calcium-dependent and exhibits higher affinity compared to known interactions.
- p30 was detected in EAT cells, mouse brain, and spleen, and purified to homogeneity.
- Partial amino acid sequencing revealed a unique protein sequence with less than 55% similarity to known proteins.
Conclusions:
- Mouse calcyclin binds to a novel protein, p30, in a calcium-dependent manner with high affinity.
- This novel protein target, p30, is expressed in various mouse tissues, suggesting a potentially widespread biological role.