Large-scale preparation of sodium-potassium ATPase from kidney outer medulla

✍ Scribed by Klodos, Irena; Esmann, Mikael; Post, Robert L.

• Publisher: Nature Publishing Group
• Year: 2002
• Tongue: English
• Weight: 74 KB
• Volume: 62
• Category: Article
• ISSN: 0085-2538
• DOI: 10.1046/j.1523-1755.2002.00654.x

✦ Synopsis

Background:

Large amounts of na,k-atpase are needed for studies involving protein chemistry. preparation of na,k-atpase from kidney by the widely used, rapid procedure of jørgensen (biochim biophys acta 356:36-52, 1974; methods enzymol 156:29-43, 1988) includes labor-intensive dissection of tissue from the outer medulla and centrifugation into a step gradient of sucrose solution.

Methods:

In a large-scale modification presented here, tissue was dissected with a surgical instrument, a rongeur, and centrifugation was simply a five times repeated differential centrifugation. the procedure took seven days and 68 person-hours of work.

Results:

The yield of activity from 26 kg of whole kidneys was 6600 units (micromol pi/min) in one preparation. the amount of protein was 240 mg and the specific activity was 28 micromol pi/min per mg protein.

Conclusions:

There is a significant saving of labor to obtain a product with a specific activity similar to that commonly obtained. the microsomal fraction may be useful for preparing other membrane proteins from the outer medulla.