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Discovery of the Nitro‐Proteome in Human Myometrium
Author(s) -
Ulrich Craig,
Nordmeier Akira,
Schegg Kathleen,
Quilici David,
Buxton Iain L. O.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.923.6
Subject(s) - myometrium , proteome , chemistry , s nitrosylation , cysteine , microbiology and biotechnology , biochemistry , biology , endocrinology , enzyme , uterus
NO nitrosylates one or more regulatory proteins in myometrial smooth muscle to produce relaxation. In order to examine nitrosylation of candidate proteins, we developed a method for showing S‐nitrosylation in whole tissue lysates. This method will allow us to elucidate the human myometrial smooth muscle nitroproteome and to test the effect of these post‐secondary modifications on relaxation. Our method, which we are calling Nitro‐DIGE selectively labels S‐nitrosylated proteins in a cell extract using spectrally resolvable Alexa Fluor maleimide dyes. Labeled extracts are then analyzed using 2‐dimensional in‐gel electrophoresis (2D DIGE). With this powerful technique we can identify differences in levels of S‐nitrosylation between term and preterm myometrium. These identified proteins are regulatory in nature and preliminary results have identified candidates such as heat shock protein 27 and alpha actinin which have been shown to regulate contraction in smooth muscle tissue. Further work is directed at both hypothesis‐directed examination of a select relaxation pathway, as well as identifying prominent proteins that are selectively nitrosylated by pregnancy progression and/or in response to experimental NO‐stimulation of tissues in functional assays where relaxation/contraction can be correlated with protein nitrosylation. Supported by March of Dimes Prematurity Initiative Grant 21‐FY10‐176 and NIH HD053028 to ILOB