Ubiquitin Modification Characterization

HEWL, penta-N-acetylchitopentaose (NAG₅), dithiotreithol (DTT), iodoacetamide, calmodulin, equine cytochrome c, horse heart myoglobin, melittin and ubiquitin were purchased from Sigma-Aldrich (Poole, UK). Trypsin and AspN were obtained from Promega (Southhampton, UK). Lys48 poly-ubiquitin ladder, wild-type USP5 and Lys48 diubiquitin were purchased from Boston Biochem (Cambridge, MA, USA). C335A USP5 was expressed and purified as previously described34 (link). Photoleucine 1 was purchased from Thermo Fisher Scientific, Loughborough, UK, and aryldiazirine 2 synthesized as described above.

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Manzi L., Barrow A.S., Scott D., Layfield R., Wright T.G., Moses J.E, & Oldham N.J. (2016). Carbene footprinting accurately maps binding sites in protein–ligand and protein–protein interactions. Nature Communications, 7, 13288.

Publication 2016

iodoacetamide calmodulin equine cytochrome c horse heart myoglobin melittin ubiquitin Trypsin AspN

Calmodulin Cytochrome c Diubiquitin Heart Horse Iodoacetamide Melittin Myoglobin Penta Poly ubiquitin Promega Trypsin Ubiquitin

Corresponding Organization :

Other organizations : University of Nottingham, Queen's Medical Centre

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Variable analysis

independent variables

Photoleucine 1
Aryldiazirine 2

dependent variables

Not explicitly mentioned

control variables

Penta-N-acetylchitopentaose (NAG5)
Dithiotreithol (DTT)
Iodoacetamide
Calmodulin
Equine cytochrome c
Horse heart myoglobin
Melittin
Ubiquitin
Trypsin
Lys48 poly-ubiquitin ladder
Wild-type USP5
Lys48 diubiquitin
C335A USP5

positive controls

Wild-type USP5
Lys48 diubiquitin

negative controls

C335A USP5

Annotations

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