Characterization and Purification

The studies of the testis LH receptor indicated that it had a molecular weight of 194000 and that it exists as a dimer with both subunits having a molecular weight of 100000 [10-13]. In contrast, photoaffinity labelling [14,15] and chemical cross-linking [16] of LH with ovarian membranes suggests that the rat ovarian receptor consists of three or four subunits. It is possible that in addition to tissue differences there are species differences in the numbers and size of the subunits, because different results have been obtained from bovine [17] and porcine ovaries [18]. 

The data available therefore indicate that the LH receptor has a molecular weight of approximately 73-100000 in the testes and the ovaries. Oligomers may exist but further confirmatory studies are required. 

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Manenti S, Sorokine O, Van Dorsseiaer A and Taniguchi H 1992 Affinity purification and characterization of myristoyiated aianine-rich protein kinase C substrate (MARCKS) from bovine brain J. Biol. Chem. 267 22 310-15... 

These methodologies have been reviewed (22). In both methods, synthesis involves assembly of protected peptide chains, deprotection, purification, and characterization. However, the soHd-phase method, pioneered by Merrifield, dominates the field of peptide chemistry (23). In SPPS, the C-terminal amino acid of the desired peptide is attached to a polymeric soHd support. The addition of amino acids (qv) requires a number of relatively simple steps that are easily automated. Therefore, SPPS contains a number of advantages compared to the solution approach, including fewer solubiUty problems, use of less specialized chemistry, potential for automation, and requirement of relatively less skilled operators (22). Additionally, intermediates are not isolated and purified, and therefore the steps can be carried out more rapidly. Moreover, the SPPS method has been shown to proceed without racemization, whereas in fragment synthesis there is always a potential for racemization. Solution synthesis provides peptides of relatively higher purity however, the addition of hplc methodologies allows for pure peptide products from SPPS as well. 

Classical gel electrophoresis has been used extensively for protein and nucleic acid purification and characterization [9, 10], but has not been used routinely for small molecule separations, other than for polypeptides. A comparison between TLC and electrophoresis reveals that while detection is usually accomplished off-line in both electrophoretic and TLC methods, the analyte remains localized in the TLC bed and the mobile phase is immediately removed subsequent to chromatographic development. In contrast, in gel electrophoresis, the gel matrix serves primarily as an anti-... 

Charbonneau, H., and Cormier, M. J. (1979). Ca2+-induced bioluminescence in Renilla reniformis. Purification and characterization of a calcium-triggered luciferin-binding protein. J. Biol. Chem. 254 769-780. 

Daubner, S. C., Astorga, A. M., Leisman, G. B., and Balwin, T. O. (1987). Yellow light emission of Vibrio barveyi strain Y-l purification and characterization of the energy-accepting yellow fluorescent protein. Proc. Natl. Acad. Sci. USA 84 8912-8916. 

Illarionov, B. A., et al. (2000). Recombinant obelin cloning and expression of cDNA, purification, and characterization as a calcium indicator. Method. Enzymol. 305 223-249. 

Kojima, S., et al. (2000a). Purification and characterization of the luciferase from the freshwater snail Latia. Abstract, 11th Int. Symp. on Biolumin. Chemilumin., Asilomar, CA, p. 57. 

Small, E. D., Koda, P., and Lee, J. (1980). Lumazine protein from the bio-luminescent bacterium Photobacterium phosphoreum. Purification and characterization./. Biol. Chem. 255 8804—8810. 

Figure 5.6 Positive-ion electrospray spectrum obtained from the major component in the LC-MS analysis of a purified recombinant 62 kDa protein using a Cig microbore 50 X 1 mm column and a flow rate of 50 p.lmin . The starting buffer (buffer A ) was 0.1% TEA in water, while the gradient buffer (buffer B ) consisted of 0.1% TEA in acetonitrile-water (9 1 vol/vol). The running conditions consisted of 0% B for 5 min, followed by a linear gradient of 100% B for 55 min. Reprinted from J. Chromatogr., B, 685, McAtee, C. P., Zhang, Y., Yarbough, P. O., Fuerst, T. R., Stone, K. L., Samander, S. and Williams, K. R., Purification and characterization of a recombinant hepatitis E protein vaccine candidate by liquid chromatography-mass spectrometry , 91-104, Copyright (1996), with permission from Elsevier Science.

Rodoni, S. et al.. Partial purification and characterization of red chlorophyll catabolite reductase, a stroma protein involved in chlorophyll breakdown. Plant Physiol, 115, 677, 1997. 

Millichip, M. et al.. Purification and characterization of oil-bodies (oleosomes) and oil-body boundary proteins (oleosins) from the developing cotyledons of sunflower (Helianthus annuus L.), Biochem. J., 314, 333, 1996. 

Bonk, M. et al.. Purification and characterization of chaperonin 60 and heat-shock protein 70 from chromoplast of Narcissus pseudonarcissus involvement of heat-shock protein 70 in a soluble protein complex containing phytoene desaturase. Plant Physiol. Ill, 931, 1996. 

Ben-Amotz, A., Kartz, A., and Avron, M., Accumulation of 3-carotene in halotolerant algae purification and characterization of 3-carotene globules from Dunaliella bardawU, J. PhycoL, 18, 529, 1983. 

Gandia-Herrero, R, Garefa-Carmona, R, and Escribano, J., Purification and characterization of a latent polyphenol oxidase from beet root Beta vulgaris L.), J. Agric. Food Chem., 52, 609, 2004. 

This paper concerns the main properties of water soluble pectins in sol and gel states. First of all, the methods of purification and characterization are discussed. The method of steric exclusion chromatography equipped with different detectors is demonstrated as the most useful to determine the macromolecular characteristics of these polymers the role of aggregation is pointed out. 

Sakamoto M, Shirane Y, Naribayashi 1, Kimura K, Morishita N, Sakamoto T, Sakai T (1994) Purification and characterization of a rhamnogalacturonase with protopectinase activity from Trametes sanguinea. EurJ Biochem 226 285-291... 

Pitkanen, K., Heikinheimo, R. and Pakkanen, R. (1992) Purification and characterization of Erwinia chrysanthemi B374 pectin methylesterase produced by Bacillus subtilis. Enzyme Microb TechnoHA, 832-836. 

P.van der Veen et al.,(1991), Induction, purification and characterization of arabinases produced by Aspergillus niger. Arch.Mcrobiol.(157)23-28. 

Pectinases from Rhizopus sp. Efficient in Enhancing the Hydrolyzation of Raw Cassava Starch Purification and Characterization... 

The present work reports the purification and characterization of acetyl esterase from orange fruit as well as the in situ localization of the enzyme by immuno histology. 

Pectin lyase from Fusarium oxysporum f. sp. radicis lycopersici purification and characterization... 

Latus M, H-J Seitz, J Eberspacher, E Lingens (1995) Purification and characterization of hydroxyquinol 1,2-dioxygenase from Azotobacter sp. strain GPl. Appl Environ Microbiol 61 2453-2460. 

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Baitsch D, C Sandn, R Brandsch, GL Igloi (2001) Gene cluster on pAOl of Arthrobacter nicotinovorans involved in degradation of the plant alkaloid nicotine cloning, purification, and characterization of 2,6-dihydroxypyridine 3-hydrolase. J Bacterid 183 5262-5267. 

Batie CJ, E LaHaie, DP Ballou (1987) Purification and characterization of phthalate oxygenase and phthalate oxygenase reductase from Pseudomonas cepacia. J Biol Chem 262 1510-1518. 

Fukumori F, RP Hausinger (1993b) Purification and characterization of 2,4-dichlorophenoxyacetate/a-keto-glutarate dioxygenase. J Biol Chem 268 24311-24317. 

Itoh N, N Morinaga, T Kouzai (1994) Purification and characterization of a novel metal-containing nonheme bromoperoxidase from Pseudomonas putida. Biochim Biophys Acta 1207 208-216. 

Kaschabek SR, T Kasberg, D Muller, AE Mars, DB Janssen, W Reineke (1998) Degradation of chloroaromat-ics purification and characterization of a novel type of chlorocatechol 2,3-dioxygenase of Pseudomonas putida GJ31. J Bacterial 180 296-302. 

Kuhm AE, A Stolz, K-L Ngai, H-J Knackmuss (1991) Purification and characterization of a 1,2-dihydroxynaphthalene dioxygenase from a bacterium that degrades naphthalenesulfonic acids. J Bacterial 173 3795-3802. 

Lehmann M, Tshisuaka B, Fetzner S, Roger P, Lingens F (1994) Purification and characterization of isoquinoline 1-oxidoreductase from Pseudomonas diminuta 7, a molybdenum-containing hydroxylase. JBiol Chem 269 11254-11260. 

Wiesner W, K-H van Pee, F Lingens (1988) Purification and characterization of a novel non-heme chloroperoxidase from Pseudomonas pyrrocinia. J Biol Chem 263 13725-13732. 

Xun L (1996) Purification and characterization of chlorophenol 4-monooxygenase from Burkholderia cepacia ACnOO. J Bacterial 178 2645-2649. 

Kengen SWM, GB Rikken, WR Hagen, CG van Ginkel, ALM Stams (1999) Purification and characterization of (per)chlorate reductase from the chlorate-respiring strain GR-1. J Bacteriol 181 6706-6711. 

Krafft T, JM Macey (1998) Purification and characterization of the respiratory arsenate reductase of Chrysio-genes arsenatis. Eur J Biochem 255 647-653. 

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