The long range objective of this project is to apply the techniques of biochemistry to investigate how genes of the nervous system are expressed and regulated. The nervous system is composed of many neurons which interconnect with one another in a precis manner. The diversity of neurons and the nature of their interactions suggest that specific genes or sets of genes are activated in some neurons and not in others. One of the specific aims of this proposal is to develop a better understanding of the molecular events associated with the expression and regulation of the most abundant and widely distributed neuropeptide the mammalian nervous system, neuropeptide Y. The specific regions of the NPY gene responsible for expression, tissue-specific expression, and response to nerve growth factor will be dissected using molecular and biochemical approaches. Most neuropeptides and numerous proteins undergo post-translational processing which requires limited proteolysis. At least two (and often more) enzymes appear to function in their processing: 1) a trypsin-like protease, which recognizes precursors to both proteins and neuropeptides where two adjacent basic residues are present; and 2) a carboxypeptidase B-like enzyme, which removes the C-terminal basic residues form the C- terminus of intermediates which are formed by limited proteolysis.
These second aim of this proposal is to better define the molecular events associated with the limited proteolysis of precursors to proteins and neuropeptides. Our laboratory has identified and sequenced a full-length clone of the carboxypeptidase B-like processing enzyme (referred to as carboxypeptidase H [CPH]). We have shown that carboxypeptidase H is synthesized as a precursor, which is the first reported example of a proteolytic-processing enzyme being synthesized as a precursor. The identification and characterization of the intermediates produced during the processing of this precursor is also a goal of this work. In addition, the gene encoding carboxypeptidase H should contain sequences which result in its selective expression in hormone producing cells. Therefore, we would like to identify and characterize these regions of the gene.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK018849-19
Application #
3226165
Study Section
Biochemistry Study Section (BIO)
Project Start
1991-10-01
Project End
1994-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
19
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Banerjee, Sourav; Ji, Chenggong; Mayfield, Joshua E et al. (2018) Ancient drug curcumin impedes 26S proteasome activity by direct inhibition of dual-specificity tyrosine-regulated kinase 2. Proc Natl Acad Sci U S A 115:8155-8160
Zhang, Hui; Zhu, Qinyu; Cui, Jixin et al. (2018) Structure and evolution of the Fam20 kinases. Nat Commun 9:1218
Qiu, Yimin; Poppleton, Erik; Mekkat, Arya et al. (2018) Enzymatic Phosphorylation of Ser in a Type I Collagen Peptide. Biophys J 115:2327-2335
Pollak, Adam J; Haghighi, Kobra; Kunduri, Swati et al. (2017) Phosphorylation of serine96 of histidine-rich calcium-binding protein by the Fam20C kinase functions to prevent cardiac arrhythmia. Proc Natl Acad Sci U S A 114:9098-9103
Cui, Jixin; Zhu, Qinyu; Zhang, Hui et al. (2017) Structure of Fam20A reveals a pseudokinase featuring a unique disulfide pattern and inverted ATP-binding. Elife 6:
Wang, Xiaorong; Cimermancic, Peter; Yu, Clinton et al. (2017) Molecular Details Underlying Dynamic Structures and Regulation of the Human 26S Proteasome. Mol Cell Proteomics 16:840-854
Nguyen, Kim B; Sreelatha, Anju; Durrant, Eric S et al. (2016) Phosphorylation of spore coat proteins by a family of atypical protein kinases. Proc Natl Acad Sci U S A 113:E3482-91
Guo, Xing; Wang, Xiaorong; Wang, Zhiping et al. (2016) Site-specific proteasome phosphorylation controls cell proliferation and tumorigenesis. Nat Cell Biol 18:202-12
Tagliabracci, Vincent S; Wiley, Sandra E; Guo, Xiao et al. (2015) A Single Kinase Generates the Majority of the Secreted Phosphoproteome. Cell 161:1619-32
He, Yantao; Guo, Xing; Yu, Zhi-Hong et al. (2015) A potent and selective inhibitor for the UBLCP1 proteasome phosphatase. Bioorg Med Chem 23:2798-809

Showing the most recent 10 out of 83 publications