The Na, K- ATPase is a member of the P-type ATPase family. It is responsible for maintaining the Na and K gradients across eukaryotic plasma membranes. The catalytic cycle of this enzyme has been described in great detail, but the events that occur on a molecular level to perform the energy-coupled ion transport are largely unknown. Crystallization of this protein, to study its structure, has been unsuccessful. To study the structure of this molecule, cysteine residues, introduced in structurally or functionally interesting positions can be probed with radioactive, fluorescent, or spin labeled reagents or be the target of site-directed cross-linking. In this study, I created one mutation using polymerase chain reaction (PCR) method. Isoleucine 788 located in the fifth transmembrane region was replaced by cysteine. This was done in a cDNA encoding for a protein devoid of cysteines in the transmembrane region. Confirmation of this mutation was done by cDNA sequencing.

• This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

Creator

• Lomino, Joseph Vincent

Publisher

• Worcester Polytechnic Institute

Identifier

• 00D137M

Advisor

• Argüello, José M.

Year

• 2000

Date created

• 2000-01-01

Resource type

• Major Qualifying Project

Major

• Biochemistry

Rights statement

• In Copyright