TY - JOUR
T1 - Linkage via K27 Bestows Ubiquitin Chains with Unique Properties among Polyubiquitins
AU - Castañeda, Carlos A.
AU - Dixon, Emma K.
AU - Walker, Olivier
AU - Chaturvedi, Apurva
AU - Nakasone, Mark A.
AU - Curtis, Joseph E.
AU - Reed, Megan R.
AU - Krueger, Susan
AU - Cropp, T. Ashton
AU - Fushman, David
N1 - Funding Information:
This work was funded by an NSF postdoctoral award to C.A.C. and NIH R01 grants GM065334 and GM021248 to D.F. and GM084396 to T.A.C.; utilized NMR instrumentation supported in part by NSF grant DBI1040158 and NIH shared instrumentation grant 1S10OD012254 , and neutron-scattering facilities supported in part by the NSF under Agreement No. DMR-0944772; and benefited from CCP-SAS software developed through a joint EPSRC (EP/K039121/1) and NSF ( CHE-1265821 ) grant. We thank Konstantin Berlin for helpful discussions regarding SES. Some mass spectrometry data were collected on a Shimadzu 8040 mass spectrometer, supported in part by a Shimadzu instrumentation award to C.A.C.
Publisher Copyright:
© 2016 Elsevier Ltd All rights reserved.
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Summary Polyubiquitination, a critical protein post-translational modification, signals for a diverse set of cellular events via the different isopeptide linkages formed between the C terminus of one ubiquitin (Ub) and the E-amine of K6, K11, K27, K29, K33, K48, or K63 of a second Ub. We assembled di-ubiquitins (Ub2) comprising every lysine linkage and examined them biochemically and structurally. Of these, K27-Ub2 is unique as it is not cleaved by most deubiquitinases. As this remains the only structurally uncharacterized lysine linkage, we comprehensively examined the structures and dynamics of K27-Ub2 using nuclear magnetic resonance, small-angle neutron scattering, and in silico ensemble modeling. Our structural data provide insights into the functional properties of K27-Ub2, in particular that K27-Ub2 may be specifically recognized by K48-selective receptor UBA2 domain from proteasomal shuttle protein hHR23a. Binding studies and mutagenesis confirmed this prediction, further highlighting structural/recognition versatility of polyubiquitins and the potential power of determining function from elucidation of conformational ensembles.
AB - Summary Polyubiquitination, a critical protein post-translational modification, signals for a diverse set of cellular events via the different isopeptide linkages formed between the C terminus of one ubiquitin (Ub) and the E-amine of K6, K11, K27, K29, K33, K48, or K63 of a second Ub. We assembled di-ubiquitins (Ub2) comprising every lysine linkage and examined them biochemically and structurally. Of these, K27-Ub2 is unique as it is not cleaved by most deubiquitinases. As this remains the only structurally uncharacterized lysine linkage, we comprehensively examined the structures and dynamics of K27-Ub2 using nuclear magnetic resonance, small-angle neutron scattering, and in silico ensemble modeling. Our structural data provide insights into the functional properties of K27-Ub2, in particular that K27-Ub2 may be specifically recognized by K48-selective receptor UBA2 domain from proteasomal shuttle protein hHR23a. Binding studies and mutagenesis confirmed this prediction, further highlighting structural/recognition versatility of polyubiquitins and the potential power of determining function from elucidation of conformational ensembles.
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U2 - 10.1016/j.str.2016.01.007
DO - 10.1016/j.str.2016.01.007
M3 - Article
C2 - 26876099
AN - SCOPUS:84959332188
SN - 0969-2126
VL - 24
SP - 423
EP - 436
JO - Structure with Folding & design
JF - Structure with Folding & design
IS - 3
ER -