Does 13C-or 15N-labeling affect Cu(I)-thiolate cluster arrangement in yeast copper-metallothionein?

It was attempted to examine whether or not isotope labeling may possibly affect an oligonuclear metal-thiolate cluster. Cu-metallothioneins are known to contain strongly distorted Cu-thiolate clusters and seemed appropriate for this study. Thus, yeast 13C-and 15N-Cu-metallothioneins were isolated from Saccharomyces cerevisiae cells grown in a minimal synthetic medium and some physicochemical parameters were compared with those of the unlabeled Cu-thionein.

Borate in mummification salts and bones from Pharaonic Egypt.

Mummification processes in Pharaonic Egypt were successful using sodium salts. Quite frequently sodium concentrations in mummified bones ranged from 300 to 4000 micromol/g. In the search for an effective inorganic conservation compound our choice fell on boric acid.

Ancient metalloenzymes as possible markers in molecular archaeology.

The successful preparation of an active remnant of Cu,Zn-superoxide dismutase from mummified brain tissue stimulated the isolation of both biochemically and immunologically active alkaline Zn2Mg-phosphatase from antique bone samples of different archaeological sites and age. In particular, specimens from pharaonic Egypt being up to 4000 years of age were used. Gel filtration, ion exchange and affinity chromatographies were employed to optimise the preparation of the ancient enzyme.

Monoclonal antibodies recognize 2300 years aged alkaline phosphatase.

It was attempted to monitor the immunological response of monoclonal antibodies directed to human alkaline phosphatase in ancient Egyptian bones from the ptolemeic period. The intactness of the respective epitopes of the bone enzyme was successfully demonstrated in an ELISA. Fortunately, the mummified bone was not contaminated by fungi and bacteria due to the fungicidal and bactericidal reactivity of the ancient pretreatment employing resins of pistachio for mummification.

Enzymatic and immunological activity of 4000 years aged bone alkaline phosphatase.

Structurally intact and functionally active human bone alkaline phosphatase was isolated from clavicle fragments of IDU, an Egyptian mummy of the Old Kingdom (2150 +/- 50 BC). Both anion exchange and affinity chromatographies were employed to optimise the preparation of the ancient enzyme resulting in a specific activity of 180 +/- 30 mU/mg. The intactness of the bone enzyme fractions of the wheat-germ lectin affinity chromatography was successfully demonstrated in an ELISA using the monoclonal antibody BAP A.

Intact mummified bone alkaline phosphatase.

Our knowledge to the mode of conservation of mummified structurally and functionally intact biopolymers is limited. Rib samples of a well-preserved 2300-year old ptolemeic mummy were examined whether or not functionally active Zn2Mg alkaline phosphatase could be detected. A protein of M(r) 170 +/- 20 kDa being close to 200 kDa of the enzyme of fresh bones was successfully isolated.

Zn2Mg alkaline phosphatase in an early ptolemeic mummy.

Bone samples of a ptolemeic mummy have been employed to study the mode of conservation on the intactness of Zn2Mg alkaline phosphatase in both structure and catalytic activity. A protein of M(r) = 190 +/- 10 kDa being identical to the 200 kDa enzyme of fresh human bones was successfully isolated. Regardless of age 200 kDa protein bands and a distinct subunit at 60 kDa were seen in SDS-PAGE electrophoresis.