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# Copyright Yair Benita Y.Benita@pharm.uu.nl
# Biopython (http://biopython.org) license applies

"""Calculate isoelectric points of polypeptides using methods of Bjellqvist.

pK values and the methos are taken from:

* Bjellqvist, B.,Hughes, G.J., Pasquali, Ch., Paquet, N., Ravier, F., Sanchez,
J.-Ch., Frutiger, S. & Hochstrasser, D.F.
The focusing positions of polypeptides in immobilized pH gradients can be predicted
from their amino acid sequences. Electrophoresis 1993, 14, 1023-1031. 

* Bjellqvist, B., Basse, B., Olsen, E. and Celis, J.E.
Reference points for comparisons of two-dimensional maps of proteins from
different human cell types defined in a pH scale where isoelectric points correlate
with polypeptide compositions. Electrophoresis 1994, 15, 529-539.

I designed the algorithm according to a note by David L. Tabb, available at:


pKinternal = {'D':4.05, 'E':4.45, 'H':5.98, 'C':9.0, 'K':10.0, 'Y':10.0, 'R':12.0}
pKcterminal= {'D':4.55, 'E':4.75}
pKnterminal = {'A':7.59, 'M':7.0, 'S':6.93, 'P':8.36, 'T':6.82, 'V':7.44, 'E':7.7}

neg = ['D', 'E', 'C', 'Y', 'Cterm']
pos = ['R', 'K', 'H', 'Nterm']

# access this module through ProtParam.ProteinAnalysis class.
# first make a ProteinAnalysis object and then call its isoelectric_point method.
class IsoelectricPoint:
      def __init__(self, ProteinSequence, AminoAcidsContent):
            self.sequence = ProteinSequence
            self.amino_acids_content = AminoAcidsContent
      #This function calculates the total charge of the protein at a given pH.            
      def _chargeR(self, pH, Cterm=0, Nterm=0):
            NegScore = 0.0
            PosScore = 0.0
            # make a local dictionary with the relevant amino acids
            for i in pKinternal.keys():
                  ChargedAminoAcids[i] = self.amino_acids_content[i]
            for i in ChargedAminoAcids.keys():
                  #first calculate the score per amino acid
                  score = ChargedAminoAcids[i] * self._cr_ratio(pKinternal[i], pH, i)
                  # determine if it goes to positive or negative
                  if i in pos:
                        PosScore += score
                  elif i in neg:
                        NegScore += score
            # now add the score for C and N terminals.
            if Nterm == 0:    
                  PosScore += self._cr_ratio(7.5, pH ,'Nterm')
                  PosScore += self._cr_ratio(Nterm, pH ,'Nterm')
            if Cterm == 0:
                  NegScore += self._cr_ratio(3.55, pH ,'Cterm')
                  NegScore += self._cr_ratio(Cterm, pH ,'Cterm')
            return PosScore-NegScore
      # This is the action function, it tries different pH until the charge of the protein is 0 (or close).
      def pi(self):           
            if self.sequence[0] in pKnterminal.keys():
                  Nterm = pKnterminal[self.sequence[0]]
                  Nterm = 0
            if self.sequence[-1] in pKcterminal.keys():
                  Cterm = pKcterminal[self.sequence[-1]]
                  Cterm = 0
            pH = 7.0
            CutBy = 3.5
            Charge = self._chargeR(pH, Cterm, Nterm)

            while CutBy > 0.0001: # this loop is usually good enough to get the job done.
                  Charge1 = self._chargeR(pH1, Cterm, Nterm)
                  Charge2 = self._chargeR(pH2, Cterm, Nterm)

                  if abs(Charge1) < abs(Charge2):
                        Charge = Charge1
                        pH = pH1
                        Charge = Charge2
                        pH = pH2
                  CutBy = CutBy/2.0
            # some times the charge is still higher or lower than zero. in such case we fix it in a step-wise way.
            if abs(Charge) > 0.1:
                  while abs(Charge) > 0.1:
                        if Charge > 0:
                              pH += 0.001
                              pH -= 0.001
                        Charge = self._chargeR(pH, Cterm, Nterm)
            return pH
      # This function calculates the charge on one amino acid at a specific pH.     
      def _cr_ratio(self, pk, ph, i):
            if i in pos:
            elif i in neg:
                  raise KeyError ("amino acid is neither positive nor negative. check dictionary.")
            return (CR/(CR+1.0))

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