PMID- 26580314
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20181202
IS  - 1860-7314 (Electronic)
IS  - 1860-6768 (Linking)
VI  - 11
IP  - 3
DP  - 2016 Mar
TI  - Pro-region engineering for improved yeast display and secretion of brain derived 
      neurotrophic factor.
PG  - 425-36
LID - 10.1002/biot.201500360 [doi]
AB  - Brain derived neurotrophic factor (BDNF) is a promising therapeutic candidate for 
      a variety of neurological diseases. However, it is difficult to produce as a 
      recombinant protein. In its native mammalian context, BDNF is first produced as a 
      pro-protein with subsequent proteolytic removal of the pro-region to yield mature 
      BDNF protein. Therefore, in an attempt to improve yeast as a host for 
      heterologous BDNF production, the BDNF pro-region was first evaluated for its 
      effects on BDNF surface display and secretion. Addition of the wild-type 
      pro-region to yeast BDNF production constructs improved BDNF folding both as a 
      surface-displayed and secreted protein in terms of binding its natural receptors 
      TrkB and p75, but titers remained low. Looking to further enhance the 
      chaperone-like functions provided by the pro-region, two rounds of directed 
      evolution were performed, yielding mutated pro-regions that further improved the 
      display and secretion properties of BDNF. Subsequent optimization of the protease 
      recognition site was used to control whether the produced protein was in pro- or 
      mature BDNF forms. Taken together, we have demonstrated an effective strategy for 
      improving BDNF compatibility with yeast protein engineering and secretion 
      platforms.
CI  - Copyright (c) 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
FAU - Burns, Michael L
AU  - Burns ML
AD  - Department of Chemical and Biological Engineering, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Malott, Thomas M
AU  - Malott TM
AD  - Department of Chemical and Biological Engineering, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Metcalf, Kevin J
AU  - Metcalf KJ
AD  - Department of Chemical and Biological Engineering, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Puguh, Arthya
AU  - Puguh A
AD  - Department of Chemical and Biological Engineering, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Chan, Jonah R
AU  - Chan JR
AD  - Department of Neurology, Program in Neuroscience, University of California, San 
      Francisco, San Francisco, California, USA.
FAU - Shusta, Eric V
AU  - Shusta EV
AD  - Department of Chemical and Biological Engineering, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA. shusta@engr.wisc.edu.
LA  - eng
GR  - P30 CA014520/CA/NCI NIH HHS/United States
GR  - T90 DK07007/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20151209
PL  - Germany
TA  - Biotechnol J
JT  - Biotechnology journal
JID - 101265833
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (NGFR protein, human)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Receptors, Nerve Growth Factor)
RN  - 0 (Recombinant Proteins)
RN  - 7171WSG8A2 (BDNF protein, human)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.1 (Receptor, trkB)
RN  - EC 2.7.10.1 (tropomyosin-related kinase-B, human)
SB  - IM
MH  - Brain-Derived Neurotrophic Factor/chemistry/*genetics/*metabolism
MH  - Membrane Glycoproteins/metabolism
MH  - Nerve Tissue Proteins/metabolism
MH  - Protein Binding
MH  - Protein Engineering/methods
MH  - Protein Folding
MH  - Protein-Tyrosine Kinases/metabolism
MH  - Receptor, trkB
MH  - Receptors, Nerve Growth Factor/metabolism
MH  - Recombinant Proteins/genetics/metabolism
MH  - Saccharomyces cerevisiae/genetics/*growth & development/metabolism
OTO - NOTNLM
OT  - Brain-derived neurotrophic factor
OT  - Directed evolution
OT  - Neurotrophin
OT  - Pro-region
OT  - Yeast surface display
EDAT- 2015/11/19 06:00
MHDA- 2016/12/15 06:00
CRDT- 2015/11/19 06:00
PHST- 2015/06/09 00:00 [received]
PHST- 2015/08/21 00:00 [revised]
PHST- 2015/10/19 00:00 [accepted]
PHST- 2015/11/19 06:00 [entrez]
PHST- 2015/11/19 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - 10.1002/biot.201500360 [doi]
PST - ppublish
SO  - Biotechnol J. 2016 Mar;11(3):425-36. doi: 10.1002/biot.201500360. Epub 2015 Dec 
      9.