Kenneth Mills

External Grants
2024-2027     National Institutes of Health, NIGMS, R15: "The Regulation of Inteins: Adaptations to Extreme Environments and Targets for Drug Discovery"

2022-2024     Research Corporation for Science Advancement, SEED (Singular Exceptional Endeavors of Discovery: "Folding Inhibitors of Inteins: a novel drug target."

2019-2023     National Institutes of Health, NIGMS, R15: "Conditional Protein Splicing of inteins from extreme thermophiles and halophiles: Structure, Mechanism, and Function."

2015-2019      National Science Foundation, RUI: "RUI: The Catalysis of Protein Splicing and the Structure and Stability of Thermophilic Inteins: An Integrated Research and Education Program"
 

2015                Research Corporation Cottrell Scholar
 

2013-2016      National Science Foundation, RUI: "Intein Structure and Catalysis of Protein Splicing and Hedgehog Autoprocessing: An Integrated Research and Education Program"

2010-2013      National Science Foundation, RUI: "Catalysis and coordination of the steps of protein splicing by non-canonical inteins: An integrated research and education program"
 

2009-2014      Henry Dreyfus Teacher-Scholar Award
 

2009                ASBMB-UAN Outreach Award
 

2005-2010      National Science Foundation, CAREER grant, “CAREER: Alternative Mechanisms of HINT domain autoprocessing: An integrated undergraduate research and education program”
 

2003-2005      National Science Foundation, Major Research Instrumentation, “Acquisition of Protein Purification and Analysis Equipment to Enhance Biochemical Research and Education at the College of the Holy Cross” 
 

2003-2005      Research Corporation, Cottrell College Science Award, “The Mechanism and Molecular Evolution of Hedgehog Proteins and Related Auto-Processing Domains”
 

2002-2004      Petroleum Research Fund (American Chemical Society), Type GB, “The Mechanism of Protein Splicing and the Molecular Evolution of Inteins.”
 

1997-2001      Howard Hughes Medical Institute Predoctoral Fellowship
 

1995-1996      Barry M. Goldwater Fellowship

(Undergraduate co-authors indicated by asterisk)

40. *Yalala, V.R., *Lynch, AK., and Mills, K.V. (2022) Conditional Alternative Protein Splicing Promoted by Inteins from Haloquadratum walsbyi.  Biochemistry, 61, 294-302.
39. *Wall, D.A., *Tarrant, S.P., Wang, C., Mills, K.V., and Lennon, C.W. (2021) Intein Inhibitors as Novel Antimicrobials: Protein Splicing in Human Pathogens, Screening Methods, and Off-Target Considerations. Front. Mol. Biosci. 8, 752824, 1-17.
38. *Williams, J.E., *Jaramillo, M.V., Li, Z., Zhao, J., Wang, C., Li, H.,  and Mills, K.V. (2021) An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein. Scientific Reports, 11, 11680, 1-12.
37. Robinzon, S., *Cawood, A.R., *Ruiz, M.A., Gophna, U., Altman-Price, N., and Mills, K.V. (2020) Protein Splicing Activity of the Haloferax volcanii PolB-c intein is sensitive to homing endonuclease domain mutations. Biochemistry, 59, 3359-3367.
36. *Chiarolanzio, K.C., *Pusztay, J.M., *Chavez, A., Zhao, J., Xie, J., Wang, C., and Mills, K.V. (2020) Allosteric Influence of Extremophile Hairpin Motif Mutations on the Protein Splicing Activity of a Hyperthermophilic Intein. Biochemistry, 59, 2459-2467.
35. Zhao, J., Du, Z., Wang, C., and Mills, K.V. (2020) Methods to Study the Structure and Catalytic Activity of cis-splicing Inteins. Methods Mol Biol, 2133, 55-73.
34. *Ramsoomair, C.K., *Yakely, A.E., *Urbanski, L.M., *Karanja, K., *Giaccone, Z.T., *Siegart, N.M., Wang, C., Gomez, A.V., Reitter, J.N., and Mills, K.V. (2017) Coordination of the third step of protein splicing in two cyanobacterial inteins. FEBS Letters, 591, 2147-2154.
33. Zwarycz, A.S., Fossat, M., Akanyeti, O., Lin, Z., Rosenman, D.J., Garcia, A., Royer, C.A., Mills, K.V., and Wang, C. (2017) V67L Mutation Fills Internal Cavity to Stabilize RecA Mtu Intein. Biochemistry 56, 2715-2722.
32.*Minteer, C.J., *Siegart, N.M., *Colelli, K.C., Liu, X., Linhardt, R.J., Wang, C., Gomez, A.O., Reitter, J.N., and Mills, K.V. (2017) Intein-promoted cyclization of aspartic acid flanking the intein leads to atypical C-terminal cleavage. Biochemistry 56, 1042-1050.
31. Reitter, J.N., *Cousin, C.E., *Nicastri, M.C., *Jaramillo, M.V., and Mills, K.V. (2016) Salt-dependent conditional protein splicing of an intein from Halobacterium salinarum. Biochemistry 55, 1279-1282.
30. Mills, K.V. (2015) Biochemistry in an undergraduate writing-intensive first-year program: Seminar courses in drugs and bioethics. Biochemistry and Molecular Biology Education 43, 263-272.
29. Kulevich, S.E., Herrick, R.S., and Mills, K.V. (2014) A Discovery Chemistry Experiment on Buffers. Journal of Chemical Education, J. Chem. Educ. 91, 1207-1211.
28. Mills, K.V., Johnson, M.A., and Perler, F.B. (2014) Protein splicing: How inteins escape from precursor proteins. Journal of Biological Chemistry, 289, 14498-14505.
27. Topilina, N.I. and Mills, K.V. (2014) Recent advances in in vivo applications of intein-mediated protein splicing. Mobile DNA 5, 5.
26. *Nicastri, M.C., *Xega, K., Li, L., Xie, J., Wang, C., Linhardt, R.J., Reitter, J.N., and Mills, K.V. (2013) Internal Disulfide Bond Acts as a Switch for Intein Activity. Biochemistry 52, 5920-5927.
25. Mills, K.V. (2013) Self-Splicing Proteins. In: Handbook of Proteolytic Enzymes, Third Edition, Vol. 1., ed. N. Rawlings and G. Salvesen, 315-321.
24. Chen, W., Li, L., Du, Z., Liu, J., Reitter, J.N., Mills, K.V., Linhardt, R.J. and Wang, C. (2012) Intramolecular Disulfide Bond between Catalytic Cysteines in an Intein Precursor. J. Am. Chem. Soc.,134, 2500-2503.
23. Du, Z., Liu, J., Albracht, C.D., Hsu, A., Chen, W., *Marieni, M.D., *Colelli, K.M., *Williams, J.E., Reitter, J.N., Mills, K.V. and Wang, C. (2011) Structural and Mutational Studies of a Hyperthermophilic Intein from DNA Polymerase II of Pyrococcus abyssi. J. Biol. Chem. 286, 38638-38648. (Cover of Nov. 4 2011 issue.)
22. Liu, J., Du, Z., Albracht, C.D., Naidu, R.O., Mills, K.V. and Wang, C. (2011) (1)H, (13)C, and (15)N NMR assignments of the Pyrococcus abyssi DNA Polymerase II intein. Biomol. NMR Assign. 5, 233-235.
21. Reitter, J.N. and Mills, K.V. (2011) Canonical protein splicing of a class one intein that has a class three non-canonical sequence motif. J. Bacteriol. 193, 994-997.
20. *O’Brien, K.M., *Schufreider, A.K., *McGill, M.A., *O’Brien, K.M., Reitter, J.N. and Mills, K.V. (2010) Mechanism of protein splicing of the Pyrococcus abyssi lon protease intein. Biochem. Biophys. Res. Commun.403, 457-461.
19. *Kerrigan, A.M, *Powers, T.L, *Dorval, D.M., Reitter, J.N. and Mills, K.V. (2009) Protein Splicing of the Three Pyrococcus abyssi ribonucleotide reductase inteins. Biochem. Biophys. Res. Commun. 387, 153-157.
18. Mills, K.V., Herrick, R.S., Ditzler, M.A., Nestor, L.P., Shafer, H., and Guilmette, L.W. (2008) Introducing Undergraduate Students to Electrochemistry: A Two-Week Discovery Chemistry Experiment. J. Chem. Educ.85, 1116-1119.
17.  Herrick, R.S., Mills, K.V., and Nestor, L.P. (2008) Data Pooling in a Chemical Kinetics Experiment: The Aquation of a Series of Cobalt (III) Complexes. J. Chem. Educ. 85, 1120-1122.
16.  Mills, K.V. and Guilmette, L.W. (2007) Thermochemical Analysis of Neutralization Reactions: An Introductory Discovery Experiment. J. Chem. Educ. 84, 326-328.
15.  Bellin, R.M. and Mills, K.V. (2007) The Holy Cross Biochemistry Concentration: An Integrated Four-Year Program to Develop Undergraduate Research Scholars. In Developing and Sustaining a Research-Supportive Curriculum, ed K.K. Karukstis and T.E. Elgren, Council on Undergraduate Research.
14.  Mills, K.V., Connor, K.R., *Dorval, D.M., and *Lewandowski, K.T. (2006) Protein Purification via Temperature-Dependent, Intein-Mediated Cleavage from an Immobilized Metal Affinity Resin.  Anal. Biochem.356, 86-93.
13.  Mills, K.V., *Dorval, D.M., *Lewandowski, K.T. (2005) Kinetic analysis of the individual steps of protein splicing for the Pyrococcus abyssi PolII intein. J. Biol. Chem 280, 2714-2720.
12. Mills, K.V., *Gareau, J.R., and *Garcia, A.M. (2005) Low-cost modification to the Copacabana method for spreading transformation mixtures.  BioTechniques39, 188.
11. Mills, K.V., Perler, F.B. (2005) The Mechanism of Intein-Based Protein Splicing: Variations on a Theme.Protein & Peptide Letters, 12, 751-755.
10. Mills, K.V. (2005) Protein Splicing. In Encyclopedia of Molecular Cell Biology and Molecular Medicine,Second Edition, ed, RA Meyers, 241-265, Weinheim: Wiley-VCH. 
9. Mills, K.V., Paulus, H. (2005) Biochemical Mechanisms of Intein-Mediated Protein Splicing. In Nucleic Acids and Molecular Biology16 (Homing Endonucleases and Inteins), ed. M. Belfort, 231-255, Berlin: Springer Verlag.
8. Mills, K.V., *Manning, J.S., *Garcia, A.M., *Wuerdeman, L.A. (2004) Protein splicing of a Pyrococcus abyssiintein with a C-terminal glutamine.  J. Biol. Chem.279, 20685-20691.
7. Mills, K.V. (2004) Self-Splicing Proteins.  In: Handbook of Proteolytic Enzymes, Second Edition, Vol. 2., ed. AJ Barrett, 2136-2140, London: Academic Press, Elsevier Science.
6. Mills, KV.  (2003) Post-Translational Control.  In: Genetics, Vol. 3., ed. R Robinson, 178-182, New York: Macmillan Reference USA.
5. Mills, K.V., Paulus, H.  (2001) Reversible Inhibition of Protein Splicing by Zinc Ion.  J. Biol. Chem. 276, 10832-10838.
4. Lew, B.M., Mills, K.V., Paulus, H. (1999) Characteristics of Protein Splicing in trans Mediated by a Semisynthetic Split Intein. Biopolymers (Peptide Science)51, 355-362.
3. Lew, B.M., Mills, K.V., Paulus, H. (1998) Protein Splicing in Vitro with a Semisynthetic Two-Component Minimal Intein. J. Biol. Chem.273, 15887-15890.
2. Mills, K.V., Lew, B.M., Jiang, Sq, Paulus, H.  (1998) Protein splicing in trans by purified N- and C- terminal fragments of the Mycobacterium tuberculosis RecA intein. Proceedings of the National Academy of Sciences, USA95, 3543-3548.
1. Broomfield, C.A., Mills, K.V., Meier, B.M., Lockridge, O.A., and Millard, C.B.  (1998) Reaction of human butyrylcholinesterase (BchE) H117 enzymes with carbamates.  In: Structure and Function of Cholinesterases and Related Proteins, 223-226.

Ken Mills, Ph.D., Principal Investigator

Libby Nompleggi, Chemistry 2025
Maggie O'Dea, Chemistry 2025
Emma Proffitt, Chemistry 2025
Caroline Quinn, Chemistry 2025

Fiona Doherty, Chemistry 2026
Alanna Mahar, Chemistry 2026
Cristina Pedorella, Chemistry/Biochem 2026

Maria Pena Garcia, Chemistry 2027
Katrin White, Biology 2027

Nana Akyeampong, Seelos STEM Scholar, 2028
Mari Arias, Seelos STEM Scholar, 2028

Seelos STEM Scholar Alumni:
Gia Ciaravino, Biology 2026
Kiara Fontes, Math 2026
Cyndie Alexis, Health Studies 2025
Maria Campos, Chemistry 2025
Brynn Dundon, Chemistry 2025

2024

Sarah Carter, Biology/Biochem, PhD Student, Dartmouth
Lilly Feeney, Chemistry 2024, Hockey, Germany (Pre Med)
Lily Nguyen, Chemistry 2024, Dental Assistant (Pre-Dental)
Kallie Poon, Biology 2024, Jesuit Volunteer Corps (Pre-Med)
Megan Yurchick, Chemistry 2024, Americorps (Pre-Med)

2023
Josue Alvarado, FRAP/Sociology 
Bea Barbesino, Chem, MD Student
Michelle Haigbea, Bio/Honors, Pharmacy student, MCPHS

2022
Justin Hackshaw, FRAP/Economics 
Anthony D'Angelo, Chem, PhD student, Texas A&M
Thuy Nguyen, Chem, Master's, Rutgers
Lily Russo, Chem, Alpha Analytical
Diana Wall, Bio, PhD student, Brown

2021
Angel Chavez, Chemistry, Pfizer
Bella Jankowski, Chem/Biochem, PhD student, UMass
Liam Lewis, Chem, Teaching
Mercedes Ruiz, English, Maxim Healthcare recruiter
Sean Tarrant, Bio/Biochem, Clinical Research Coordinator
Vaish Yalala, Chemistry, MD Student, UMass
 
2020
Alexandra Cawood, Chem/Biochem, DO Student, W. Virginia SOM
Chinnel Hudson, FRAP/Sociology

 2019
Owen Conroy, Biology, Broad Institute
Patrick Exconde, Chem/Biochem, PhD student UPenn
Abby Lynch, Biology/Biochem, Research Assistant Duke
Nhi Tran, UMass Amherst
Amanda Wibben, Biology Fulbright India, MS Theology Harvard, MD Student Georgetown

 2018
Hannah Comeau, Chemistry, Masters, Johns Hopkins, Clinical Research Coordinator
Vanessa Costa, Sociology, Clinical specialist
C.J. Janton, Bio/Biochem, DO/PhD student, NYIT
Christian Ramsoomair, Chem/Biochem MD/PhD student, Miami
Deirdre Reidy, Biology, MD student, UConn

2017
Shirley Amunya, Fitchburg St, Teach for America
Alexander Bonano, Spanish, MPH, Tufts
Josh Long, Biology, MD 
Chris Minteer, Chem/Biochem PhD, Yale
Wanlin Zhang, Chem/Biochem, DDS, Touro

2016
Chris Cousin, Chemistry, PharmD, MCPHS
Julliard del Rosario, Cornell U
Mike Koulopoulos, Chem/Biochem, MD Wake Forest
Anthony Yakely, Chem/Biochem, JVC Northwest, MPH Yale

  2015
Dawilmer Castillo, Bridgewater St., Masters, Brown, Pharmacist
Zach Giaccone, Chem/Biochem, Dental School, UConn
Mario Jaramillo, Biology, MD, Albany Med
Isabelle Pierre, UMass Dartmouth
Nikki Siegart, Bio/Biochem, MD SUNY Downstate

  2014
Sarah Cahn, Chemistry, Takeda Oncology
Dan Haley, Chemistry DDS, UConn

  2013
Katie Colelli Chiarolanzio, Bio/Biochem, M.D., UVM
Nick Gulbrandsen, Bio Fitchburg
Michael Nicastri, Chem/Biochem PhD, UPenn, PD Columbia
Jen Pusztay, Chem/Biochem , D.O. UNECOM
Laura Urbanski , Chem/Biochem MD/PhD program, UConn
Jennie Williams, Chemistry , Program Director, Lovelane Riding

  2012
Stacy Chin, Chemistry, PhD, Boston U
Kevin Karanja, Chemistry WSU, Science Teacher
George Savidis, Bio/Biochem, Cytivia

  2011
Colleen Donahue, Biology, M.D., Tufts
Michelle Marieni, Biology, M.D., Creighton
Kevin O’Brien, Bio/Biochem PhD, U. Georgia, PD UMass Med
Kristina Xega, Chemistry, MS Harvard, Charles River

  2010
Lauren Duffee, Bio/Biochem, D.V.M., UPenn
Adam Nadelson, Biology, M.D., UMass
Dan York, Bio/Biochem, MD, Texas Tech

  2009
Adam Kerrigan, Biology Cybereason
Annie Schufreider, Chemistry, M.D., Loyola Chicago
Jen Winslow, Biology, DO, University of New England

  2008
Melissa McGill, Bio/Biochem, M.S., Michigan, Pfizer
Kathryn O’Brien, Chem/Biochem PhD, Cornell-Weill

  2007
Matt Drago, Bio/Biochem, M.D., UPenn
Taryn Powers Surabian, Biology Ursuline Academy

  2006
Deirdre Dorval Wells, Biology J.D., Harvard Law School

  2005
Katie Lewandowski Bagnowski, Bio/Biochem PhD, Wash. U., PD Northwestern

  2004
Kate Leahy, Math, MD, Boston U
Jackie Gareau Merrick, Chem/Biochem, Ph.D, Cornell-Weill
Tim Quinn, Chem/Biochem, M.D.-Georgetown

  2003
Jenny Manning Chapnick, Biology
Lisa Wuerdeman Lucas, Chemistry D.O., U. New England
Ralph Millilo, Bio/Biochem, M.D. SUNY Upstate
Lisa Garcia Seals, Biology Middle School Teacher

  2002
Beth Ambrose Dean, Chemistry Physician Assistant
Pat Twomey, Chem/Biochem, M.D., Georgetown

Kate Connor, Research Assistant
Alvin Gomez, Research Associate 
Julie Reitter, Ph.D., Research Associate 

Our lab studies protein splicing and the enzymology of extreme thermophiles.

Protein splicing is a post-translational modification. It involves the self-catalyzed excision of an intervening polypeptide sequence, the intein, concomitant to the ligation of the flanking polypeptides, the exteins, via a native peptide bond.

Below is a schematic of the protein splicing process:

We are interested in

a) The means by which non-canonical inteins facilitate the individual steps of protein splicing.

b) The means by which inteins coordinate the steps of splicing.

c) The role that inteins play in the control of extein function.

d) Why are inteins there? What is their potential role in the organism?

e) Comparisons of the catalytic strategies of enzymes from halophiles, thermophiles, and other extremophiles and their mesophilic equivalents.

Below is the standard chemical mechanism of protein splicing: