Chloroplast Proteins Extraction

The chloroplast proteome plays a crucial role in the entire cellular metabolism, including photosynthesis and amino acid biosynthesis. The preparation of complete and purified maize chloroplasts is an important basis for the study of maize chloroplast proteomics, photosynthesis and its regulatory mechanism. Lifeasible's expertise in chloroplast engineering enables us to obtain purified, intact chloroplast proteins from a wide variety of plants. If you have any special requirements about our chloroplast proteins extraction service, please feel free to contact us.

Chloroplast Proteins

Chloroplasts are important organelles for photosynthesis in higher plants. Contains a large amount of protein in chloroplasts. Studies have shown that the whole proteome in higher plants contains about 21000-25000 proteins, and the proteins in chloroplasts account for 10%-25% of the whole plant tissue proteins. This shows that chloroplasts play a very important role in plant cells. The chloroplast proteome and subproteome include chloroplast outer and inner envelope, soluble matrix in chloroplast, thylakoid membrane and thylakoid lumen. In recent years, comparative proteomic analysis of chloroplasts by 2-DE has received extensive attention. The application of 2-DE to the entire chloroplast proteome remains challenging due to the complex composition of chloroplast proteins, especially the membrane system. Therefore, it is necessary to establish a system suitable for the isolation of various plant chloroplasts and their proteome research, which will lay the foundation for further research on chloroplast proteomics.

Fig. 1. Theoretical enrichment factor (EF) of envelope proteins, from crude cell extract to purified envelope fractions. (Takamatsu T, et al., 2018)

Services

In recent years, there have been many improvements in chloroplast protein extraction methods for different types of plants. The preparation of high-quality chloroplast proteins is an important basis for the study of plant chloroplast proteomics, photosynthesis and its regulatory mechanism. Lifeasible is committed to providing high-quality chloroplast protein extraction services to customers around the world. Optimized for chloroplast proteome yield, purity and minimal protein degradation, our scientists applied different protein extraction procedures to purified chloroplasts. We have successfully extracted purified and complete chloroplast protein from sugarcane, spinach, corn, Arabidopsis, rice and other plants. In addition, we used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization mass spectrometry (ESI-MS) to identify single proteins after protein separation. The basic process of chloroplast protein extraction is as follows:

(1) Intact chloroplasts were prepared by Percoll gradient centrifugation or sucrose density gradient centrifugation.

(2) Determination of chlorophyll content.

(3) Chloroplast integrity was detected by light microscopy and chloroplast envelope integrity was determined by the Hill reaction rate.

(4) Chloroplast proteins were extracted and determined by TCA acetone precipitation method, Trizol precipitation method, Tris-HCl method, urea-thiourea method and phenol method.

(5) SDS-PAGE and two-dimensional electrophoresis.

Competitive Advantages

• The extraction efficiency of chloroplast protein is high, and the amount of protein is large.
• Selecting the appropriate protein extraction program system according to the needs.
• Advanced separation techniques are characterized by high sensitivity and low sample volume.
• Multiple samples can be analyzed simultaneously, reducing workload and cost.
• Rich experience in chloroplast protein extraction.

References

1. Fan P, Wang X, Kuang T, et al. (2009) An efficient method for the extraction of chloroplast proteins compatible for 2‐DE and MS analysis[J]. Electrophoresis. 30(17): 3024-3033.
2. Bouchnak I, Brugière S, Moyet L, et al. (2019) Unraveling Hidden Components of the Chloroplast Envelope Proteome: Opportunities and Limits of Better MS Sensitivity*[S][J]. Molecular & Cellular Proteomics. 18(7): 1285-1306.