pi = (pk 1 + pk 2 )/2, the ph where the molecule has a net charge of 0

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pI = (pk 1 + pk 2 )/2, the pH where the molecule has a net charge of 0. Ninhydrin Test. Ion Exchange Chromatography. Step 1. Separation of Polypeptide Chains. dissociated by exposure to pH extremes, 8 M urea, 6 M guanidinium hydrochloride, or high salt cencentrations. - PowerPoint PPT Presentation

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pI = (pk1 + pk2)/2, the pH where the molecule has a net charge of 0

Phe-Ser-Tyr-Gly-Val-Gln

64 69

O2N

N

NO

HH

Phe

O

Val

OGln

HO

blue

green

green fluorescent protein (GFP)

Ninhydrin Test

Ion Exchange Chromatography

Protein separation

size exclusion chromatography

ultrafiltration

ultracentrifugation

least soluble at their IP

Step 1. Separation of Polypeptide Chains

dissociated by exposure to pH extremes, 8 M urea, 6 M guanidinium hydrochloride, or high salt cencentrations

Cleavage of disulfide bridges

Edman degradation

phenylisothiocyanate

50 cycles of reaction can be accomplished on 50 pmol (about 0.1 g)

Carboxypeptidases cleave amino acid residues from the C-termini

Cyanogen bromide

Fmoc solid phase peptide synthesis: A Practical Approach

Edited by

Weng C. Chan and Peter D. White

OXFORD UNIVERSITYPRESS

2000