Related Experiment Video
Updated: Jul 17, 2026
11:32
Determination of the Relative Cell Surface and Total Expression of Recombinant Ion Channels Using Flow Cytometry
Published on: September 28, 2016
[Recombinant expression of mutant human annexin V]
Jiong Cai1, Fang L I, Ying-Qiang Zhang
1Department of Nuclear Medicine, PUMC Hospital, CAMS and PUMC, Beijing 100730, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. Acta Academiae Medicinae Sinicae
|September 26, 2006
Summary
Researchers successfully expressed highly active mutant human annexin V in Pichia Pastoris. This recombinant protein possesses endogenous metal-chelating sites, offering potential for new therapeutic applications.
Area of Science:
- Recombinant protein expression
- Molecular biology
- Biochemistry
Context:
- Annexin V is a calcium-dependent phospholipid-binding protein involved in membrane trafficking and apoptosis.
- Developing recombinant annexin V with enhanced functional properties is of significant interest for therapeutic and diagnostic applications.
Purpose:
- To engineer and express a mutant human annexin V with an endogenous metal-chelating site in the Pichia Pastoris expression system.
- To achieve purification of active mutant human annexin V.
Summary:
- The human annexin V gene was mutated at the 5' and 3' ends and cloned into the pPIC9K vector for Pichia Pastoris transformation.
- Transformed Pichia Pastoris secreted a 36 kDa protein upon methanol induction, exhibiting high binding activity.
- The mutant annexin V demonstrated potent inhibition of fluorescein-annexin V binding to phosphatidylserine-exposed erythrocytes.
Impact:
- Successful expression of highly active recombinant mutant human annexin V in Pichia Pastoris.
- The engineered protein possesses endogenous metal-chelating sites, expanding its potential applications.
- This study provides a robust system for producing functional annexin V variants.
