Re: [ccp4bb] Sequence of crystallised protein fragment

- Protein crystallography

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: CNS error message with MR
From: Christian Biertuempfel biertumpfelc {- at -} NIDDK {- dot -} NIH {- dot -} GOV
Date: 2008-07-02

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Subject: crystallization tips for acidic proteins
From: "Karakas, Erkan" karakas {- at -} CSHL {- dot -} EDU
Date: 2008-07-02

Subject: Re: Sequence of crystallised protein fragment
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-07-02

Tryptic digest of the excised gel band, followed by rigorous peptide
matching helped us identify a fragment in a very recent case. The
crystallographic results confirmed MS findings.

Curiously, the ends were 'ragged' so direct ESI-MS was not very useful,
likewise the N-terminal sequencing would not have been a good idea because
of the heterogenous N-terminus.

Artem

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Klaus
Futterer
Sent: Tuesday, July 01, 2008 7:11 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Sequence of crystallised protein fragment

We have a 150 kDa protein that reproducibly crystallises at one of
the Hampton Screen conditions. However, we know from SDS gel analysis
that the crystals contain only a 45 kDa fragment, that forms through
proteolytic cleavage over time. We would like to determine the
sequence boundaries of the fragment.

We believe a combination of N-terminal sequencing plus MS analysis
might give us the information we need, but I was wondering whether
the ccp4bb community has other suggestions.

Klaus

---------------------------------------------------------------------

Klaus Fütterer, Ph.D.

School of Biosciences P: +44-(0)-121-414 5895
University of Birmingham F: +44-(0)-121-414 5925
Edgbaston E: k.futterer@bham.ac.uk
Birmingham, B15 2TT, UK W: www.biochemistry.bham.ac.uk/klaus/
---------------------------------------------------------------------

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: CNS error message with MR
From: Christian Biertuempfel biertumpfelc {- at -} NIDDK {- dot -} NIH {- dot -} GOV
Date: 2008-07-02

Next message:
Subject: crystallization tips for acidic proteins
From: "Karakas, Erkan" karakas {- at -} CSHL {- dot -} EDU
Date: 2008-07-02