135:. It is a dipteran-specific toxin and it plays an important role in how to produce a bioinsecticide to control mosquitoes. So, it is very essential to determine which functional motif of Cry4Aa contributes to this activity. In this study, several Cry4Aa mutants were made by replacing the residues of potential receptor binding site, loops 1, 2, and 3 in domain II with alanine. A bioassay
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kalata B1. Cyclotides display a wide range of pharmaceutically important bioactivities, but their natural function is in plant defense as insecticidal agents. On the structure of cyclotides kalata B1, all 23 non-cysteine residues were successively substituted with alanine. The data were tested by
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Alanine scanning was used to determine simultaneously the functional contributions of 19 side chains buried at the interface between human growth hormone and the extracellular domain of its receptor. Each amino acid in the side chains was substituted by alanine. Then shotgun scanning method which
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This technique is rapid, because many side chains are analyzed simultaneously and the need for protein purification and biophysical analysis is circumvented. The technology is very mature at this point and is widely used in biochemical fields. The data can be tested by
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One good example of alanine scanning is the examination of the role of charged residues on the surface of proteins. In a systematic study on the roles of conserved charged residues on the surface of epithelial sodium channel
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Edelheit O, Hanukoglu I, Dascal N, Hanukoglu A (April 2011). "Identification of the roles of conserved charged residues in the extracellular domain of an epithelial sodium channel (ENaC) subunit by alanine mutagenesis".
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of mutated protein remains intact, as Ala mimics the secondary structure preferences of the majority of the encoded or canonical amino acids. This is predicted by the
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Another critical application of alanine scanning is to determine the influence of individual residues on structure and activity in the prototypic
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In addition, alanine scanning is also used to determine which functional motif of Cry4Aa has the mosquitocidal activity. Cry4Aa was produced by
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101:), alanine scanning was used to reveal the importance of charged residues for the process of transport of the proteins to the cell surface.
81:. Furthermore, computational methods to estimate thermodynamic parameters based on theoretical alanine substitutions have been developed.
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324:"Alanine scanning analyses of the three major loops in domain II of Bacillus thuringiensis mosquitocidal toxin Cry4Aa"
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Example of alanine scanning. The native protein (top row) and each possible point mutation to alanine is considered.
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technique used to determine the contribution of a specific residue to the stability or function of a given protein.
280:"Alanine scanning mutagenesis of the prototypic cyclotide reveals a cluster of residues essential for bioactivity"
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is used because of its non-bulky, chemically inert, methyl functional group that nevertheless mimics the
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Howlader MT, Kagawa Y, Miyakawa A, Yamamoto A, Taniguchi T, Hayakawa T, Sakai H (February 2010).
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preferences that many of the other amino acids possess. Sometimes bulky amino acids such as
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Gauguin L, Delaine C, Alvino CL, McNeil KA, Wallace JC, Forbes BE, De Meyts P (July 2008).
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Simonsen SM, Sando L, Rosengren KJ, Wang CK, Colgrave ML, Daly NL, Craik DJ (April 2008).
381:"Alanine scanning of a putative receptor binding surface of insulin-like growth factor-I"
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218:"Rapid mapping of protein functional epitopes by combinatorial alanine scanning"
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are used in cases where conservation of the size of mutated residues is needed.
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Morrison KL, Weiss GA (June 2001). "Combinatorial alanine-scanning".
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Weiss GA, Watanabe CK, Zhong A, Goddard A, Sidhu SS (August 2000).
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The alanine scanning method takes advantage of the fact that most
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Genetic techniques for biological research: a case study approach
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and binomial mutagenesis with phage display technology was used.
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This technique can also be used to determine whether the
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can be exchanged with Ala by point mutations, while the
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110:combines the concepts of alanine scanning
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141:was followed to test the activities.
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70:. This is usually accomplished by
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486:. London: J. Wiley. p. 111.
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436:Am. J. Physiol. Renal Physiol
192:10.1016/S1367-5931(00)00206-4
512:Molecular biology techniques
222:Proc. Natl. Acad. Sci. U.S.A
66:plays a significant role in
448:10.1152/ajprenal.00648.2010
16:Molecular biology technique
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74:or randomly by creating a
72:site-directed mutagenesis
37:site-directed mutagenesis
328:Appl. Environ. Microbiol
398:10.1074/jbc.M802620200
297:10.1074/jbc.M709303200
243:10.1073/pnas.160252097
132:Bacillus thuringiensis
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151:canonical amino acids
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348:10.1128/AEM.02175-09
482:Michels CA (2002).
340:2010ApEnM..76..860H
234:2000PNAS...97.8950W
180:Curr Opin Chem Biol
159:Alanine-World model
155:secondary structure
145:Alanine-World model
45:secondary structure
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493:978-0-471-89921-1
29:molecular biology
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124:NMR Spectroscopy
91:NMR Spectroscopy
33:alanine scanning
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475:Further reading
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62:of a specific
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105:Applications
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112:mutagenesis
68:bioactivity
165:References
60:side chain
119:cyclotide
506:Category
456:21209000
417:18502759
366:19948851
306:18258598
262:10908667
200:11479122
408:3258947
357:2813026
336:Bibcode
230:Bibcode
79:library
64:residue
53:leucine
41:Alanine
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464:869654
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49:valine
460:S2CID
253:16802
35:is a
488:ISBN
452:PMID
413:PMID
362:PMID
302:PMID
258:PMID
196:PMID
99:ENaC
444:doi
440:300
403:PMC
393:doi
389:283
352:PMC
344:doi
292:doi
288:283
248:PMC
238:doi
188:doi
76:PCR
51:or
27:In
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