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Assignment on Dornase Alpha PDF

Added on - 27 Jul 2022

BIOLOGY. BIOLOGY. 7. 7. : BIOLOGY. DORNASE ALPHA. Name

Pet Protein Study: Sequence and Purification for DORNASE ALFA You will be given the name of a protein of industrial/medical importance. You will need to select a specific species variant that is used in a medical or industrial application. Find out about: 1. The primary structure (amino acid sequence) of your Pet Protein. Prepare your own figure (not a journal extract) of the primary sequence, clearly showing the numbering relevant to the mature sequence. Outline and label any sub-regions or domain units important in this protein. Indicate on this figure the disulfide connections (if any), as well as any amino acids known to be glycosylated. 2. The physico-chemical properties (pI, MW value) of the protein that are important considerations for a purification protocol. Use the tools in ExPASy to analyse your sequence. Check you have the mature sequence of your protein (i.e. remove any signal peptide) If a glycoprotein, detail the full chemical nature of the carbohydrate por

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Running head: BIOLOGY
DORNASE ALPHA
Name of the Student
Name of the University
Author Note
BIOLOGY1
Table of Contents
Introduction................................................................................................................................2
1. The primary structure (amino acid sequence):.......................................................................2
2. The physico-chemical properties (pI, MW value) of the protein that are important
considerations for a purification protocol:.................................................................................0
Predicted proteins..................................................................................................0
ProtScale..................................................................................................................................1
User-provided sequence:...................................................................................................1
3. How the protein was initially purified from its natural source when it was first discovered:3
4. How is the same protein now prepared for an industrial use, or in recombinant form:.........4
Bibliography...............................................................................................................................6
BIOLOGY2
Introduction
Dornase alfa is a highly purified solution containing the recombinant DNase I protein of
human beings. This is an enzyme solution which is responsible for the selective cleavage of
DNA. This enzyme has been found to hydrolyse the DNA which has been found to be present
in the sputum/mucus of the patients suffering from cystic fibrosis and has been found to
reduce viscosity in lungs in order to facilitate airway clearance. The trade name of this
enzyme is Pulmozyme. This paper will discuss the primary structure, physicochemical
properties and purification processes of Dornase alfa.
1. The primary structure (amino acid sequence):
1020304050
MRGMKLLGALLALAALLQGAVSLKIAAFNIQTFGETKMSNATLVSYIVQI
60708090100
LSRYDIALVQEVRDSHLTAVGKLLDNLNQDAPDTYHYVVSEPLGRNSYKE
110120130140150
RYLFVYRPDQVSAVDSYYYDDGCEPCGNDTFNREPAIVRFFSRFTEVREF
160170180190200
AIVPLHAAPGDAVAEIDALYDVYLDVQEKWGLEDVMLMGDFNAGCSYVRP
210220230240250
SQWSSIRLWTSPTFQWLIPDSADTTATPTHCAYDRIVVAGMLLRGAVVPD
260270280
SALPFNFQAAYGLSDQLAQAISDHYPVEVMLK
Signal peptide sequence= Positions 1 to 22 [Yellow]
Mature chain sequence= Positions 23 to 282 [Green]
Glycosylation= N-linked glycosylation of 40thresidue the protein (Asparagine) [Red] and N-
linked glycosylation of 128thresidue the protein (Asparagine) [Red].
Disulphide bond= 123rdto 136thresidue and 195thto 231stresidue. The latter one is
responsible for enzyme activities
BIOLOGY3
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