CLOSTRIPAIN Property

storage temp. 

2-8°C

form 

lyophilized powder

color 

White to off-white

Water Solubility 

Soluble in water

Specific Activity

≥20units/mg solid

Safety

Safety Statements 

22-24/25

WGK Germany 

3

F 

3-10-21

Hazard and Precautionary Statements (GHS)

Symbol(GHS)

Signal word

Danger

Hazard statements

H315Causes skin irritation

H319Causes serious eye irritation

H334May cause allergy or asthma symptoms or breathing difficulties if inhaled

H335May cause respiratory irritation

Precautionary statements

P261Avoid breathing dust/fume/gas/mist/vapours/spray.

P264Wash hands thoroughly after handling.

P264Wash skin thouroughly after handling.

P271Use only outdoors or in a well-ventilated area.

P302+P352IF ON SKIN: wash with plenty of soap and water.

P305+P351+P338IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continuerinsing.

CLOSTRIPAIN Chemical Properties,Usage,Production

Uses

A protease that cleaves proteins on the carboxyl bond of arginine

Uses

Clostripain from Clostridium histolyticum has been used as a proteolytic enzyme in perfusate to detect its effect on tube hematocrit. It has also been used in limited proteolysis of DNA polymerase (gp43) of phage T4 (RB69 gp43).

Biochem/physiol Actions

Clostripain from Clostridium histolyticum is composed of two polypeptide chains, with molecular masses of 41.7 kDa and 15.4 kDa. Clostripain has a highly restricted substrate specificity for Arg-Xaa peptide bonds. Therefore, clostripain has been explored as a potential enzyme for protein sequencing purposes. It has also been studied as a catalyst for condensation of pharmaceutically important peptides containing Arg-Pro bonds.

Purification Methods

Clostripain is isolated from Clostridium histolyticum callogenase by extraction in pH 6.7 buffer, followed by hydroxylapatite chromatography with a 0.1-0.2 M phosphate gradient, then Sephadex G-75 gel filtration with 0.05M phosphate pH 6.7, dialysis and a second hydroxylapatite chromatography (gradient elution with 0.1M 0.3M phosphate, pH 6.7). It has proteinase and esterase activity and is assayed by hydrolysing N-benzoyl-L-arginine methyl ester. [Mitchell & Harrington J Biol Chem 243 4683 1968, Methods Enzymol 19 635 1970.]