164610 Sigma-Aldrichp-Aminophenylmercuric Acetate - CAS 6283-24-5 - Calbiochem

Safety Data Sheet (SDS) 

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision	06-September-2007 RFH
Synonyms	APMA
Description	An organomercurial agent used for the activation of latent matrix metalloproteinases (MMPs) in vitro. Acts by facilitating the loss of the enzyme propeptide domain through an autolytic cleavage known as the cysteine switch.
Form	Yellow solid
Recommended reaction conditions	The MMPs are members of a family of at least twenty Zn2+-dependent endopeptidases that function extracellularly at neutral pH. The MMPs are initially synthesized as latent or inactive zymogens that contain a regulatory propeptide. This propeptide confers the latency by binding to the Zn2+ at the enzyme's active site. MMP activation can be achieved by autoactivation and by the action of other MMPs or proteases, such as furin, urokinase, plasmin, and trypsin, as well as by organomercurial compounds. The following is a protocol from Stricklin, et al. (1983. Biochemistry 22, 61) describing the use of paminophenylmercuric acetate (APMA) to activate pro-MMPs. This protocol is good for other types of organomercurials such as p-(hydroxymercuric) benzoate (PHMB), phenylmercuric chloride (PMC), and mersalyl. Organomercurial stock solutions Prepare stock solutions in 0.1 M NaOH at a concentration range of 10-50 mM just prior to use. Although not absolutely necessary, the stock solution may be adjusted to pH 11 with 5 N HCl. (e.g. Marcy, A.I. et al. 1991. Biochemistry 30, 6476). ProMMPs Prepare proenzyme solutions in 0.1 M Tris-HCl, pH 7.5 at 1-2 mg/ml. Activation of proMMPs To initiate activation, combine proMMP solution with organomercurial stock in a 10:1 volume* ratio (MMPs to organomercurial). Incubate the mixture at 37°C for 2-3 hrs**. *When more organomercurial is desired, make the stock solution more concentrated. Do not exceed the above ratio so that the volume of organomercurial in the mixture is more than about 1/10 of that of the proMMP solution; this could result in significant changes in pH. **It is recommended that an analytical run be conducted first to determine the optimal incubation time. For example, a small-scale experiment with fixed concentration of proMMP and organomercurial would be incubated as described above. Remove aliquots of sample at various time points. Stop the reaction by the addition of concentrated SDS-sample buffer (for example, add 10 µl of 2X sample buffer to a 10 µl aliquot or add 2 µl of 5X sample buffer to an 8 µl aliquot) and then boil. The progress of the activation can then be monitored qualitatively by analyzing these time aliquots on a 12% SDS-PAGE gel. Removing organomercurial from the mixture after activation The activated MMPs can be used without the removal of the organomercurial. Researchers are referred to the following reference for the removal of organomercurial by gel filtration if desired: Marcy, A.I., et al. 1991. Biochemistry 30, 6476
CAS number	6283-24-5
RTECS	OV5550000
Chemical formula	C₈H₉HgNO₂
Structure formula
Purity	≥90% by titration
Solubility	DMSO (10 mg/ml) or 0.1 N NaOH (10 mg/ml)
Storage	+15°C to +30°C
Do Not Freeze	Ok to freeze
Special Instructions	Following reconstitution, refrigerate (4°C). Stock solutions are stable for up to 1 month at 4°C.
Toxicity	Highly Toxic
References	Chen, J.-M., et al. 1991. J. Biol. Chem. 266, 5113. Marcy, A.I., et al. 1991. Biochemistry 30, 6476. Stricklin, G.P., et al. 1983. Biochemistry 22, 61.