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. Author manuscript; available in PMC: 2012 Nov 9.
Published in final edited form as: J Am Chem Soc. 2011 Oct 19;133(44):17855–17868. doi: 10.1021/ja207220g

Scheme 3.

Scheme 3

Schematics on the assembly of long Ub chains with either homogeneous or mixed linkages. Shown in (a,b) are two methods to form a homogeneously K11-linked Ub4 from either (a) two K11-linked dimers, or (b) a combination of a homogeneously K11-linked Ub3 and K11Boc Ub monomer. (c) The ability to make Ub chains with mixed linkages can be easily exploited with our present method. Here we illustrate the formation of a mixed K33,K11-linked Ub3 from a K33-linked Ub2 and K11Boc Ub monomer. The dotted line indicates the isopeptide bond that would be formed during the ligation reaction. Longer Ub chains can be assembled iteratively by using the same concepts as illustrated here.