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Expression of recombinant proteins lacking methionine as N-terminal amino acid in plastids: Human serum albumin as a case study

Paper ID Volume ID Publish Year Pages File Format Full-Text
25509 43578 2007 12 PDF Available
Title
Expression of recombinant proteins lacking methionine as N-terminal amino acid in plastids: Human serum albumin as a case study
Abstract

Removal of the N-terminal methionine of a protein could be critical for its function and stability. Post-translational modifications of recombinant proteins expressed in heterologous systems may change amino-terminal regions. We studied the expression of mature proteins lacking methionine as the N-terminal amino acid in tobacco chloroplasts, using human serum albumin (HSA) as an example. Two approaches were explored. First, we fused the Rubisco small subunit transit peptide to HSA. This chimeric protein was correctly processed in the stroma of the chloroplast and rendered the mature HSA. The second approach took advantage of the endogenous N-terminal methionine cleavage by methionine aminopeptidase. Study of this protein processing reveals a systematic cleavage rule depending on the size of the second amino acid. Analysis of several foreign proteins expressed in tobacco chloroplasts showed a cleavage pattern in accordance to that of endogenous proteins. This knowledge should be taken into account when recombinant proteins with N-terminus relevant for its function are expressed in plastids.

Keywords
Plastid transformation; Human serum albumin; Methionine removal; N-end rule; Post-translational modifications; Transit peptide
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Expression of recombinant proteins lacking methionine as N-terminal amino acid in plastids: Human serum albumin as a case study
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 4, 20 January 2007, Pages 593–604
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us