From: Richard.PedrettiAllen@octel.com
  To  : sjlessey@concentric.net
  Date: Thu, 31 Jul 1997 13:13:38 -0700

RE: Denaturing Protein with Gua-HCl

Well, if you're talking about unconventional serine proteases, of
course, everyone knows you've got to be more careful.

-Richard

p.s. I am so tired of playing this "Witness Protection" game... I want
to be back in the lab!!


>{*} Not really since you don't have much proteolytic activity in 6M
>{*} guanidinium at any temperature, since it is such a strong 
>{*} chaotropic agent.
>{*} 
>
>One has to approach this with caution.  It depends on the type of 
>protease you are using.  There are a few proteases that are fully
>active in 6 M Gdn.HCl or 9 M Urea.  If using the conventional 
>serine proteases then Richards' statement would be o.k.
>
>Cheers
>-raman
>___________________________________________________________________
>C.S.Raman                    Tel: (714) 824-4322
>University of California     Fax: (714) 824-8540 
>Dept. MB & B                 email: raman@indigo1.bio.uci.edu
>3205 Bio Sci II
>Irvine, CA 92697-3900
>-------------------------------------------------------------------
>In a time of drastic change it is the learners who inherit the 
>future.  The learned usually find themselves equipped to live
>in a world that no longer exists.  --Eric Hoffe
>___________________________________________________________
>
>My advice is an obvious oversimplification of a relatively trivial
>question but I'm not about to go into protracted dissertations about
>protein decomposition or quantifications of reaction times in a series
>of graded temperature classes with a complete stranger!  My goodness!
>What would people think?!  
>
>Besides, this a distinguished member of the Department of Biochemical &
>Molecular Biology of Oklahoma State University!  Why doesn't he go to
>the library and look it up?
>
>Mind the gap.
>
>Cheers, Richard
>
>>----------
>>From: 	ANDREWS, CHRIS[SMTP:ANDREWSC@pab281a.ssd.loral.com]
>>Sent: 	Thursday, July 31, 1997 11:36 AM
>>To: 	Davis, Michael via voicemail; 72650.3277; ANDREWS.CHRIS; bill.thomson;
>>catfishzz; dbrenner; dky; droo.thomson; dsteinberg; gluthr; grenier2;
>>kamradtJ; ospivey; paulb; paulg; RASMB; Richard.PedrettiAllen; russell;
>>sjlessey; wsand
>>Subject: 	RE: Denaturing Protein with Gua-HCl
>>
>>
>>Well ,  I must say that I agree with Richard's assessment.
>>After all , you can't make butter with a toothpick , you know what
>>I'm sayin'?
>> ----------
>>From: Richard.PedrettiAllen
>>To: Michael.Davis; 72650.3277; ANDREWS.CHRIS; bill.thomson; catfishzz; 
>>dbrenner; dky; droo.thomson; gluthr; grenier2; kamradtJ; paulb; paulg; 
>>richard.pedrettiallen; russell; sjlessey; wsand; dsteinberg; ospivey; RASMB
>>Subject: RE: Denaturing Protein with Gua-HCl
>>Date: Thursday, July 31, 1997 10:14AM
>>
>>Not really since you don't have much proteolytic activity in 6M
>>guanidinium at any temperature, since it is such a strong
>>chaotropic agent.
>>
>>Cheers, Richard
>>>
>>>Dear RASMBers,
>>>
>>>   When testing for protein subunit molec. weights using 6 M
>>>guanidinium-HCl, is it wise to add the protein sample to nearly boiling
>>>guanidinium-HCl to avoid potential proteolytic activities as is done with
>>>SDS denaturation?
>>>
>>>   Thanks,
>>>   Olin
>>>
>>>H. Olin Spivey                    Phone: (405) 744-6192 or -6189
>>>Dept of Biochem. & Molec. Biol.     Fax: (405) 744-7799
>>>246 NRC
>>>Oklahoma State University
>>>Stillwater, OK 74078-0454        E-Mail: Ospivey@BIOCHEM.OKSTATE.EDU
>>>
>>>
>>>(See attached file: rfc822.txt)
>>>
>>
>