1. My recentresearchfocuseson3 directions:
(1) One focus of our research is on the structures and functions of fibrinolytic proteins,
including urokinase (uPA), its receptor (uPAR), inhibitor (PAI-1), and tPA [1-3]. We
determined the crystal structures of uPAR [4-6], uPAR:vitronectin complex [7], uPA in
complex withvariousinhibitors,and the Michael complexesof tPA:PAI-1[8] anduPA:PAI-1
[9].PAI-1anduPA are biomarkersforcancermetastasis, havingthe highestlevelof clinical
evidence,levelof evidence 1,asrecommendedby AmericanSocietyforClinical Oncology
andthe GermanBreastCancer. However,noinhibitorsof eitheruPAor PAI-1gointoclinical
trials. We developed inhibitorsfor PAI-1 (small molecule [10] and protein-based, named
PAItrap[11]) anduPA (cyclicpeptides)[12-15],anddiscovered newprinciplesforinhibitor
development[12,14, 16]. The studywas extendedtoothertypesof proteases,including
kallikrein [17],matriptase[18,19],andfactorXI.We alsodevelopedinhibitorsforuPAR [2,
20], and one inhibitor (ATF) was fused to human serum albumin (HSA) by recombinant
technique tonotonly increase plasmahalf-life,butalsotorenderdrug carrierproperties
of the fusionprotein [21,22].
(2) Anotherfocusison structuresof cell surface receptors [18, 19, 23-25] and humanserum
albumin [26-32]. Despite its long history, HSA remains to be an actively studied protein
due to its wide applications and its abundance and important functions in human (~0.4
mMin humanblood).Ideterminedaseriesof crystal structuresof HSA inthe presence of
variousligands.A surprising findingisthe conversionof glucose fromcyclicformtolinear
forminducedbyHSA asdefinedbyourcrystal structurestudies[30],demonstratinganew
functionforthisoldprotein[29].
(3) On the area of chemical biology, we developed a technology for biomedical imaging,
cancertherapy,andantimicrobial applications.We synthesizedauniquefluorescentprobe
(beta-CarboxyZincPhthalocyanine,CPZ[33]) inlarge quantity,which hasthree important
features: (a) it has strong fluorescence (ex610nm/em690nm); (b) when illuminated at
680nm, CPZgeneratesreactive oxygenatedspecies(ROS);(c)CPZcontainsacarboxygroup
which can be activated and attached to protein or peptide. We conjugated CPZ to a
number of peptides: (a) urokinase receptor targeting peptide (ATF, [34]); (b) polylysine
peptide [35, 36]; (c) Gonadotropin-releasinghormone (GnRH) peptide targetingatGnRH
receptor[37],and demonstratedthe tumortargetingcapabilityof the conjugatesinmice
models. We alsousedthe lysine-conjugatedCPZforantimicrobialapplications [38-40] and
demonstrated that it also inactivated drug resistant bacterial strains.This conjugate was
later found practical applications in facial masks and various fabric materials
(http://www.sundynamictech.com/).
References:
See also
https://www.ncbi.nlm.nih.gov/pubmed/?term=Huang%2CMingdong%5BAU%5D+OR+Huang
%2CMing-dong%5BAU%5D
1. Yuan, C. and M. Huang, Does the urokinase receptor exist in a latent form? Cell Mol
Life Sci,2007. 64(9): p. 1033-7.

2. 2. Chen, Z., et al., Challenges for drug discovery - a case study of urokinase receptor
inhibition. CombChemHigh ThroughputScreen,2009. 12(10): p. 961-7.
3. Ngo,J.C.,etal., Structuralbasisfortherapeuticintervention of uPA/uPARsystem. Curr
Drug Targets,2011. 12(12): p. 1729-43.
4. Huai, Q., et al., Structure of human urokinaseplasminogen activatorin complex with
its receptor. Science,2006. 311(5761): p.656-9.
5. Xu, X., et al., Identification of a new epitope in uPAR as a target for the cancer
therapeuticmonoclonalantibody ATN-658,a structuralhomolog of the uPAR binding
integrin CD11b (alphaM). PLoSOne,2014. 9(1): p. e85349.
6. Lin, L., et al., Structure-based engineering of species selectivity in the interaction
between urokinase and its receptor: implication for preclinical cancer therapy. J Biol
Chem,2010. 285(14): p.10982-92.
7. Huai, Q., et al., Crystalstructures of two human vitronectin,urokinaseand urokinase
receptorcomplexes. NatStructMol Biol,2008. 15(4): p. 422-3.
8. Gong, L., et al., Crystal Structure of the Michaelis Complex between Tissue-type
Plasminogen Activator and Plasminogen Activators Inhibitor-1. J Biol Chem, 2015.
290(43): p. 25795-804.
9. Lin,Z., et al., Structuralbasisfor recognition of urokinase-typeplasminogen activator
by plasminogen activatorinhibitor-1. JBiol Chem, 2011. 286(9): p.7027-32.
10. Lin, Z., et al., Structural insight into inactivation of plasminogen activator inhibitor-1
by a small-moleculeantagonist. ChemBiol,2013. 20(2): p. 253-61.
11. Gong, L., et al., Structural basis of specific inhibition of tissue-type plasminogen
activatorby plasminogen activatorsinhibitor-1.DataBrief,2016. 6: p.550-5.
12. Jiang, L., et al., Distinctive binding modes and inhibitory mechanisms of two peptidic
inhibitors of urokinase-type plasminogen activator with isomeric P1 residues. Int J
BiochemCell Biol,2015. 62: p. 88-92.
13. Sorensen,H.P.,etal.,Selection of High-Affinity PeptidicSerineProteaseInhibitorswith
Increased Binding Entropy froma Back-Flip Library of Peptide-ProteaseFusions. JMol
Biol,2015. 427(19): p.3110-22.
14. Jiang, L., et al., Insights into the serine protease mechanism based on structural
observations of the conversion of a peptidyl serine protease inhibitor to a substrate.
BiochimBiophysActa,2016. 1860(3): p.599-606.
15. Kromann-Hansen,T.,etal.,A Camelid-derived AntibodyFragmentTargeting theActive
Site of a Serine Protease Balances between Inhibitor and Substrate Behavior. J Biol
Chem,2016. 291(29): p.15156-68.
16. Jiang,L.,etal., Rezymogenationof activeurokinaseinduced by an inhibitory antibody.
BiochemJ,2013. 449(1): p. 161-6.
17. Xu,P., etal., Design of Specific Serine ProteaseInhibitorsBased on a Versatile Peptide
Scaffold: Conversion of a Urokinase Inhibitor to a Plasma Kallikrein Inhibitor. J Med
Chem,2015. 58(22): p. 8868-76.
18. Yuan, C.,et al., Structureof catalyticdomain of Matriptasein complex with Sunflower
trypsin inhibitor-1.BMC Struct Biol,2011. 11: p. 30.
19. Zhao,B.,etal., Crystalstructuresof matriptasein complex with itsinhibitorhepatocyte
growthfactoractivatorinhibitor-1.JBiol Chem, 2013. 288(16): p. 11155-64.

3. 20. Rullo, A.F., et al., Re-engineering the Immune Response to Metastatic Cancer:
Antibody-Recruiting SmallMolecules Targeting the UrokinaseReceptor. Angew Chem
Int EdEngl, 2016. 55(11): p. 3642-6.
21. Li, R., et al., A novel tumor targeting drug carrier for optical imaging and therapy.
Theranostics,2014. 4(6): p. 642-59.
22. Zheng,K.,et al., Dual actionsof albumin packaging and tumortargeting enhancethe
antitumor efficacy and reduce the cardiotoxicity of doxorubicin in vivo. Int J
Nanomedicine,2015. 10: p. 5327-42.
23. Yuan, C., et al., Crystal structures of the ligand-binding region of uPARAP: effect of
calcium ion binding. BiochemJ,2016. 473(15): p. 2359-68.
24. Jiang, L., et al., Dimer conformation of soluble PECAM-1, an endothelial marker. Int J
BiochemCell Biol,2016. 77(Pt A):p. 102-8.
25. Liu,M., et al., Recombinanthepatocytegrowth factoractivatorinhibitor1:expression
in Drosophila S2 cells, purification and crystallization. Acta Crystallogr F Struct Biol
Commun,2017. 73(Pt 1): p.45-50.
26. Guo, S., et al., Structural basis of transport of lysophospholipids by human serum
albumin. BiochemJ,2009. 423(1): p. 23-30.
27. Luo, Z., et al., Structural evidence of perfluorooctane sulfonate transport by human
serumalbumin. ChemResToxicol,2012. 25(5): p. 990-2.
28. Wang, Y., et al., A fluorescent fatty acid probe, DAUDA, selectively displaces two
myristatesbound in human serumalbumin. ProteinSci,2011. 20(12): p.2095-101.
29. Wang, Y., S. Wang, and M. Huang, Structureand enzymaticactivities of human serum
albumin. Curr PharmDes,2015. 21(14): p. 1831-6.
30. Wang, Y., et al., Structural mechanism of ring-opening reaction of glucose by human
serumalbumin. J Biol Chem,2013. 288(22): p. 15980-7.
31. Yang,F.,etal., Effectofhuman serumalbumin on drugmetabolism:structuralevidence
of esteraseactivity of human serumalbumin. JStruct Biol,2007. 157(2): p. 348-55.
32. Zhu, L., et al., A new drug binding subsite on human serum albumin and drug-drug
interaction studied by X-ray crystallography. JStructBiol,2008. 162(1): p. 40-9.
33. Chen,J.,etal., Derivatizablephthalocyaninewith singlecarboxylgroup:Synthesisand
purification. InorganicChemistryCommunications,2006. 9(3): p. 313-315.
34. Chen,Z., et al., Zinc phthalocyanineconjugated with theamino-terminalfragmentof
urokinasefortumor-targeting photodynamictherapy. ActaBiomater,2014. 10(10): p.
4257-68.
35. Chen, Z., et al., Pentalysine beta-carbonylphthalocyanine zinc: an effective tumor-
targeting photosensitizer for photodynamic therapy. ChemMedChem, 2010. 5(6): p.
890-8.
36. Li,L., etal., Enhanced photodynamicefficacy of zincphthalocyanineby conjugating to
heptalysine. BioconjugChem,2012. 23(11): p. 2168-72.
37. Xu, P., et al., Receptor-targeting phthalocyanine photosensitizer for improving
antitumorphotocytotoxicity. PLoSOne,2012. 7(5): p. e37051.
38. Chen,J.,etal., Substituted zincphthalocyanineasan antimicrobialphotosensitizerfor
periodontitis treatment. Journal of Porphyrins and Phthalocyanines,2011. 15(04): p.
293-299.

4. 39. Chen,Z., etal., Photodynamicantimicrobialchemotherapy using zincphthalocyanine
derivativesin treatmentof bacterialskin infection.JBiomedOpt,2016. 21(1): p.18001.
40. Chen, Z., et al., An effective zinc phthalocyanine derivative for photodynamic
antimicrobialchemotherapy. Journal of Luminescence,2014. 152: p. 103-107.