Detection of Phospho-Tyrosine Hydroxylase (S31) by Western Blot Western blot of PC-12 cells incubated in the absence (Control) and presence of okadaic acid (OA, 1 μM for 60 minutes) showing specific immunolabeling of the approximately 62 kDa Tyrosine Hydroxylase phosphorylated at S31.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
For long-term storage, ≤ -20° C is recommended. Product is stable at ≤ -20° C for at least 1 year.
Background: Tyrosine Hydroxylase
Rat tyrosine hydroxylase (TH) is a pterin-dependent monooxygenase that catalyzes the hydroxylation of tyrosine to DOPA through the use of a non-heme iron. It is a 55 kDa, 498 amino acid (aa) alpha -helical protein that runs anomalously at 62 kDa in SDS-PAGE. The molecule contains a 164 aa N-terminal regulatory region and a 334 aa C-terminal catalytic domain. Its activity is regulated at the post-transcriptional level by phosphorylation of serine and feedback by catecholamines. Four serines are known to be phosphorylated at aa positions 8, 19, 31, and 40. Different kinases contribute to different phosphorylation patterns. For example, MAPKAPK-2 and CaMKII act on S19 and S40, PKA phosphorylates only at S40, while Cdk5 phosphorylates S31. Variable site phosphorylations have variable effects. S40 phosphorylation blocks catecholamine feedback inhibition. Subsequent phosphorylations at S19 or S31 likely stabilize an otherwise unstable enzyme phosphorylated only at S40, contributing to increased enzyme activity. Phosphorylation at S8 is likely to be physiologically unimportant.
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