Developments in Structural Genomics: Protein Purification and Function Interpretation

2004 ◽  
Vol 5 (1) ◽  
pp. 37-48 ◽  
Author(s):  
Cong-Zhao Zhou ◽  
Yu-Xing Chen

Proteins are an integral part of molecular and cellular structure and function and are probably the most purified type of biological molecule. In order to elucidate the structure and function of any protein it is first necessary to purify it. Protein purification techniques have evolved over the past ten years with improvements in equipment control, automation, and separation materials, and the introduction of new techniques such as affinity membranes and expanded beds. These developments have reduced the workload involved in protein purification, but there is still a need to consider how unit operations linked together to form a purification strategy, which can be scaled up if necessary. The two Practical Approach books on protein purification have therefore been thoroughly updated and rewritten where necessary. The core of both books is the provision of detailed practical guidelines aimed particularly at laboratory scale purification. Information on scale-up considerations is given where appropriate. The books are not comprehensive but do cover the major laboratory techniques and common sources of protein. Protein Purification Techniques focuses on unit operations and analytical techniques. It starts with an overview of purification strategy and then covers initial extraction and clarification techniques. The rest of the book concentrates on different purification methods with the emphasis being on chromatography. The final chapter considers general scale-up considerations. Protein Purification Applications describes purification strategies from common sources: mammalian cell culture, microbial cell culture, milk, animal tissue, and plant tissue. It also includes chapters on purification of inclusion bodies, fusion proteins, and purification for crystallography. A purification strategy that can produce a highly pure single protein from a crude mixture of proteins, carbohydrates, lipids, and cell debris to is a work of art to be admired. These books (available individually or as a set)are designed to give the laboratory worker the information needed to undertake the challenge of designing such a strategy.


2001 ◽  
Vol 123 (19) ◽  
pp. 4619-4620 ◽  
Author(s):  
Pinghua Liu ◽  
Kazuo Murakami ◽  
Takayuki Seki ◽  
Xuemei He ◽  
Siu-Man Yeung ◽  
...  

1978 ◽  
Vol 33 (1-2) ◽  
pp. 108-112 ◽  
Author(s):  
Salil Bose ◽  
George E. Hodi

Abstract 9-Aminoacridine (9AA) binds to photosynthetic m em branes of unilluminated chloroplasts in low-salt media. The binding was insensitive to the uncouplers of photophosphorylation. The apparent binding constant was 140 μM . The binding isotherm as a function of 9AA concentration was sigmoid, and approximately 3 mol 9AA/mol chlorophyll was bound at saturating concentrations of 9AA.Addition of Mg2+ partially reversed the binding of 9AA in chloroplasts in the dark as observed by a Mg2+-induced increase of 9AA fluorescence as well as by spectrophotom etric measurements of free 9AA. It appeared, however, that use of fluorescence techniques for m easuring free 9AA introduced an error in the estimation of the m agnitude of binding, particularly at low concentration of 9AA ( < 7 5 μM). This is probably due to change in fluorescence yield of membrane-bound 9AA on addition of cations. The nature of the binding of 9AA to the thylakoid membranes and the effects of Mg2+ thereupon suggest that both chemical binding of cations and screening of surface charge of the mem branes should be considered in discussing the mechanism of cation action on chloroplast structure and function. Interpretation of these data with respect to heterogeneity of sites of cation action upon or within chloroplast membranes is discussed.


Genomics could be viewed as the study of the randomness of DNA sequences. It may be possible to predict the structure of a gene product from the nucleotide sequences and thereby predict its function. The terms “structural genomics” and “functional genomics” were coined to denote the assignment of structure and function to a gene product, respectively. Proteomics focuses on the products of gene, which are basically proteins. Proteins are responsible for the development of phenotype, and proteomics is the bridge between genotype and phenotype. The transcribed mRNAs and their abundance are called transcriptome. Proteomics also deals with the interaction between proteins called intractomics. Metabolomics is concerned with identification, abundance, and localization of all the molecules excluding lipids and polysaccharides in the cell. In this chapter, the basic concepts and analysis of the genomic, proteomic, and metabolomics data for their practical utilization are discussed.


2004 ◽  
Vol 5 (1/2) ◽  
pp. 111-118 ◽  
Author(s):  
Youngchang Kim ◽  
Irina Dementieva ◽  
Min Zhou ◽  
Ruiying Wu ◽  
Lour Lezondra ◽  
...  

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