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Title
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Crystallization and preliminary X-ray analysis of human transglutaminase 3 from zymogen to active form.
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Authors
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H.C.Kim,
Z.Nemes,
W.W.Idler,
C.C.Hyde,
P.M.Steinert,
B.Ahvazi.
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Ref.
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J Struct Biol, 2001,
135,
73-77.
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PubMed id
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Abstract
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Transglutaminases(TGases; protein-glutamine-glutamyl-transferases) are a large
family of calcium-dependent acyl-transfer enzymes that catalyze the formation of
covalent cross links in proteins. Of these, the "epidermal" or "hair follicle"
TGase 3 isoform is critically involved in barrier formation in epithelia. It is
a zymogen, requiring proteolytic activation to achieve maximal specific
activity. In order to understand its structure and function, we have devised
methods for the rapid large-scale expression of the TGase 3 zymogen in the
baculovirus system, and here we describe the purification of the zymogen and
activated forms. We describe methods for the formation of high-quality,
well-diffracting crystals within 3-5 days, using both dioxane and
beta-octylglucoside to overcome severe twinning problems. The crystal of the
zymogen belongs to the triclinic space group P1 and diffracts to 2.2-A
resolution, and the crystal of the active form belongs to the P2(1) space group
at 2.7-A resolution.
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